Functional analysis of metal tolerance proteins isolated from Zn/Cd hyperaccumulating ecotype and non-hyperaccumulating ecotype of <i>Sedum alfredii</i>
 Hance

Zhang, Min; Senoura, Takeshi; Yang, Xiaoe; Nishizawa, Naoko K.

doi:10.1016/j.febslet.2011.07.013

Cited by 43 publications

(25 citation statements)

References 31 publications

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“…Besides the different stability ranking resulted from distinct algorithms, another discrepancy was observed in the stability ranking with four kinds of heavy metals treatments using same software, which might imply that diverse reactions appeared in Hyperaccumulator S. alfredii in response to different heavy metals stresses. In previous studies, ACT was often used as internal control for expression quantification of heavy metals stresses responsive genes in S. alfredii [20-22]. In our study, ACT was also regarded as stable reference genes, however, UBC9 and TUB were suggested to be better for RT-qPCR normalization in S. alfredii , which might provide more accurate expression measurement of target genes.…”

Section: Discussionmentioning

confidence: 73%

“…The ideal reference genes should be expressed at a relatively constant level among samples from different tissues and developmental stages, and also should be unaffected by different experimental treatments [17-19]. During the previous gene expression analysis in S. alfredii , ACT was often used for RT-qPCR normalization [20-22], however, without any systematic study validating reference genes under abiotic stress reported for S. alfredii . Recent studies revealed that some traditional housekeeping genes such as ACT , TUB and EF-1α were not constantly stably expressed under different experimental conditions [23-25].…”

Section: Introductionmentioning

confidence: 99%

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Selection and Validation of Reference Genes for Real-Time Quantitative PCR in Hyperaccumulating Ecotype of Sedum alfredii under Different Heavy Metals Stresses

et al. 2013

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Real-time Quantitative PCR (RT-qPCR) has become an effective method for accurate analysis of gene expression in several biological systems as well as under different experimental conditions. Although with high sensitivity, specificity and broad dynamic range, this method requires suitable reference genes for transcript normalization in order to guarantee reproducible and meaningful results. In the present study, we evaluated five traditional housekeeping genes and five novel reference genes in Hyperaccumulating ecotype of Sedum alfredii, a well known hyperaccumulator for heavy metals phytoremediation, under Cd, Pb, Zn and Cu stresses of seven different durations. The expression stability of these ten candidates were determined with three programs - geNorm, NormFinder and BestKeeper. The results showed that all the selected reference genes except for SAND could be used for RT-qPCR normalization. Among them UBC9 and TUB were ranked as the most stable candidates across all samples by three programs together. For the least stable reference genes, however, BestKeeper produced different results compared with geNorm and NormFinder. Meanwhile, the expression profiles of PCS under Cd, Pb, Zn and Cu stresses were assessed using UBC9 and TUB respectively, and similar trends were obtained from the results of the two groups. The distinct expression patterns of PCS indicated that various strategies could be taken by plants in adaption to different heavy metals stresses. This study will provide appropriate reference genes for further gene expression quantification using RT-qPCR in Hyperaccumulator S. alfredii.

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Section: Discussionmentioning

confidence: 73%

Section: Introductionmentioning

confidence: 99%

Selection and Validation of Reference Genes for Real-Time Quantitative PCR in Hyperaccumulating Ecotype of Sedum alfredii under Different Heavy Metals Stresses

et al. 2013

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“…MTP1 genes from the Zn/Cd hyperaccumulating and non-hyperaccumulating ecotypes of Sedum alfredii were both able to suppress Zn hypersensitivity in the zrc1 yeast mutant (Table 1), induced Zn accumulation and were localized at the tonoplasts of onion cells (Zhang et al, 2011). As described for A. halleri (Dräger et al, 2004), the amount of MTP1 transcripts of the hyperaccumulating ecotype is higher in shoots than in roots, a difference not observed in the non-hyperaccumulator ecotype.…”

Section: Mtp Proteins In Metal Hyperaccumulator Speciesmentioning

confidence: 99%

“…As described for A. halleri (Dräger et al, 2004), the amount of MTP1 transcripts of the hyperaccumulating ecotype is higher in shoots than in roots, a difference not observed in the non-hyperaccumulator ecotype. Upon Zn or Cd excess treatment, MTP1 transcripts accumulate in shoots of the hyperaccumulator and in roots of the non-hyperaccumulator ecotype (Zhang et al, 2011). These results indicate that, as in other hyperaccumulator species, S. alfredii hypertolerance probably relies on increased expression of MTP1 transporters in shoots.…”

Section: Mtp Proteins In Metal Hyperaccumulator Speciesmentioning

confidence: 99%

Roles of plant metal tolerance proteins (MTP) in metal storage and potential use in biofortification strategies

Ricachenevsky¹,

Menguer²,

Sperotto³

et al. 2013

Front. Plant Sci.

159

118

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Zinc (Zn) is an essential micronutrient for plants, playing catalytic or structural roles in enzymes, transcription factors, ribosomes, and membranes. In humans, Zn deficiency is the second most common mineral nutritional disorder, affecting around 30% of the world's population. People living in poverty usually have diets based on milled cereals, which contain low Zn concentrations. Biofortification of crops is an attractive cost-effective solution for low mineral dietary intake. In order to increase the amounts of bioavailable Zn in crop edible portions, it is necessary to understand how plants take up, distribute, and store Zn within their tissues, as well as to characterize potential candidate genes for biotechnological manipulation. The metal tolerance proteins (MTP) were described as metal efflux transporters from the cytoplasm, transporting mainly Zn2+ but also Mn2+, Fe2+, Cd2+, Co2+, and Ni2+. Substrate specificity appears to be conserved in phylogenetically related proteins. MTPs characterized so far in plants have a role in general Zn homeostasis and tolerance to Zn excess; in tolerance to excess Mn and also in the response to iron (Fe) deficiency. More recently, the first MTPs in crop species have been functionally characterized. In Zn hyperaccumulator plants, the MTP1 protein is related to hypertolerance to elevated Zn concentrations. Here, we review the current knowledge on this protein family, as well as biochemical functions and physiological roles of MTP transporters in Zn hyperaccumulators and non-accumulators. The potential applications of MTP transporters in biofortification efforts are discussed.

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“…MTP1 is a key transporter modulating the VSC of Zn (Sinclair and Krämer, 2012; Ricachenevsky et al, 2013). Piled evidences suggest that the expression level of MTP1 is greatly enhanced in various metal hyperaccumulators (Assunção et al, 2001; Persans et al, 2001; Becher et al, 2004; Dräger et al, 2004; Hammond et al, 2006; Talke et al, 2006; van de Mortel et al, 2006; Gustin et al, 2009; Shahzad et al, 2010; Zhang et al, 2011), which later on was shown to be attributable to genome amplification of gene copies, in contrast to a single copy in their relative non-hyperaccumulators (Dräger et al, 2004; Shahzad et al, 2010). A similar regulation of TcHMA3 (Heavy Metal ATPases 3) was also observed (Ueno et al, 2011), among which gene amplification leads to much higher expression of TcHMA3 in the accumulating ecotype Ganges of Thlaspi caerulescens than in the non-accumulating Prayon, thus VSC of Cd in roots is greatly reduced and more metal is driven to translocate to shoots.…”

Section: Regulation Of Vscmentioning

confidence: 99%

Vacuolar sequestration capacity and long-distance metal transport in plants

Peng

Gong

2014

Front. Plant Sci.

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The vacuole is a pivotal organelle functioning in storage of metabolites, mineral nutrients, and toxicants in higher plants. Accumulating evidence indicates that in addition to its storage role, the vacuole contributes essentially to long-distance transport of metals, through the modulation of Vacuolar sequestration capacity (VSC) which is shown to be primarily controlled by cytosolic metal chelators and tonoplast-localized transporters, or the interaction between them. Plants adapt to their environments by dynamic regulation of VSC for specific metals and hence targeting metals to specific tissues. Study of VSC provides not only a new angle to understand the long-distance root-to-shoot transport of minerals in plants, but also an efficient way to biofortify essential mineral nutrients or to phytoremediate non-essential metal pollution. The current review will focus on the most recent proceedings on the interaction mechanisms between VSC regulation and long-distance metal transport.

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Functional analysis of metal tolerance proteins isolated from Zn/Cd hyperaccumulating ecotype and non-hyperaccumulating ecotype of Sedum alfredii Hance

Abstract: a b s t r a c tThe Zn/Cd hyperaccumulating ecotype (HE)

Cited by 43 publications

References 31 publications

Selection and Validation of Reference Genes for Real-Time Quantitative PCR in Hyperaccumulating Ecotype of Sedum alfredii under Different Heavy Metals Stresses

Selection and Validation of Reference Genes for Real-Time Quantitative PCR in Hyperaccumulating Ecotype of Sedum alfredii under Different Heavy Metals Stresses

Roles of plant metal tolerance proteins (MTP) in metal storage and potential use in biofortification strategies

Vacuolar sequestration capacity and long-distance metal transport in plants

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