Lhcb2 gene expression analysis in two ecotypes of Sedum alfredii subjected to Zn/Cd treatments with functional analysis of SaLhcb2 isolated from a Zn/Cd hyperaccumulator

MOE Key Lab of Environmental Remediation and Ecosystem Health, College of Environmental & Resource Sciences, Zhejiang University, Hangzhou, Zhejiang 310029, China.
Biotechnology Letters (Impact Factor: 1.59). 04/2011; 33(9):1865-71. DOI: 10.1007/s10529-011-0622-8
Source: PubMed


The Lhcb2 gene from hyperaccumulator Sedum alfredii was up-regulated more than three-fold while the non-hyperaccumulator accumulated one or two-fold higher amount of the mRNA than control plants under different concentrations of Cd(2+) for 24 h. Lhcb2 expression was up-regulated more than five-fold in a non-hyperaccumulator S. alfredii when exposed to 2 μM Cd(2+) or 50 μM Zn(2+) for 8 d and the hyperaccumulator had over two-fold more mRNA abundance than the control plants. Over-expression of SaLhcb2 increased the shoot biomass by 14-41% and the root biomass by 21-57% without Cd(2+) treatment. Four transgenic tobacco lines (L5, L7, L10 and L11) possessed higher shoot biomass than WT plants with Cd(2+). Four transgenic lines (L7, L8, L10 and L11) accumulated 6-35% higher Cd(2+) amounts in shoots than the wild type plants.

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    • "Gene specific primers (Table 1) were designed using the software Primer Premier 5.0. The conditions and primers for real-time PCR were first optimized and these PCR products were sequenced for confirmation as detailed in our previous study (Yueen Chao et al., 2010; Zhang et al., 2011a, 2011b). In brief, reactions were carried out using the SYBR Green premix Ex Taq kit (Takara, DaLian, China) on a Mastercycler ep realplex2 (Eppendorf, Hamburg, Germany). "
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    ABSTRACT: Sedum alfredii Hance is a promising cadmium (Cd) hyperaccumulating plant recently identified in China. However, the physiological and molecular mechanisms underlying Cd accumulation, which differentiate hyperaccumulating ecotype (HE) from non-hyperaccumulating ecotype (NHE) has not been elucidated yet. A hydroponic experiment was conducted to investigate the role of sulfur assimilation pathway in Cd hyperaccumulation by the S. alfredii Hance, by analyzing gene expression pattern in sulfur assimilation pathway and the concentration of some sulfur containing compounds. The results show that, sulfur assimilation pathway was affected by Cd differently in HE and NHE S. alfredii Hance. The gene expression pattern of sulfur assimilation pathway was regulated differently in HE and NHE plants, especially the nicotianamine synthase (NAS). NAS transcript levels in root of HE was 141-fold higher than NHE, while in shoots of HE only 0.31-fold higher than NHE. In HE roots, NAS expression level was maximum 3171-fold higher than shoots, while in NHE plants roots NAS expression level was maximum 45.3-fold higher than shoots. In HE plant roots, sulfur, cysteine and methionine concentrations increased 30%, 46% and 835% respectively, by Cd treatment, but in NHE plants roots, sulfur concentration increased less than 1%, cysteine and methionine concentrations decreased 78.5% and 13.3% respectively, by Cd. Cd exposure increased glutathione levels by 142% in HE but less than 10% in NHE plant roots.
    Full-text · Article · Nov 2013 · Ecotoxicology and Environmental Safety
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    • "Differential level of expression in Lhcb2 was observed in case of two ecotypes of Sedum (hyperaccumulating and non-hyperaccumulating) which suggested temporal regulation of gene expression. Upon 24 hrs of Cd (2μ M) treatment non-hyperaccumulating ecotype exhibited higher expression level than hyperaccumulating, followed by a reversal of the situation after 8 days (Zhang et al. 2011). "
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    ABSTRACT: Cadmium is a potentially toxic heavy metal that hampers plant productivity by interfering with their photochemistry. Cd causes disturbances in a range of physiological processes of plants such as photosynthesis, water relations, ion metabolism and mineral uptake. Cd pronouncedly affects photosynthesis by alteration of its vital machinery in all aspects. Photosynthesis is a well organised and sequential process fundamental to all green plants and microorganisms which involves various components, including photosynthetic pigments and photosystems, the electron transport system and CO2 reduction pathways. Any damage at any level caused by Cd, critically affects overall photosynthetic capacity. Present review focuses on key effects of Cd on photosynthetic apparatus including chloroplast structure, photosynthetic pigments, Chl-protein complexes and photosystems resulting in overall decrease in efficiency of carbon assimilation pathway.
    Full-text · Article · Oct 2013
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    ABSTRACT: 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.
    Full-text · Article · Dec 2013 · PLoS ONE
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