Heavy metal pollution of agricultural soils is one of the most severe ecological problems in the world. Prosopis juliflora, a phreatophytic tree species, grows well in heavy metal laden industrial sites and is known to accumulate heavy metals. Heavy Metal ATPases (HMAs) are ATP driven heavy metal pumps that translocate heavy metals across biological membranes thus helping the plant in heavy metal tolerance and phytoremediation. In the present study we have isolated and characterized a novel 28.9 kDa heavy metal ATPase peptide (PjHMT) from P. juliflora which shows high similarity to the C-terminal region of P ATPase HMA1. It also shows the absence of the invariant signature sequence DKTGT, and the metal binding CPX motif but the presence of conserved regions like MVGEGINDAPAL (ATP binding consensus sequence), HEGGTLLVCLNS (metal binding domain) and MLTGD, GEGIND and HEGG motifs which play important roles in metal transport or ATP binding. PjHMT, was found to be upregulated under cadmium and zinc stress. Heterologous expression of PjHMT in yeast showed a higher accumulation and tolerance of heavy metals in yeast. Further, transgenic tobacco plants constitutively expressing PjHMT also showed increased accumulation and tolerance to cadmium. Thus, this study suggests that the transport peptide from P. juliflora may have an important role in Cd uptake and thus in phytoremediation.
Metallothioneins are involved in detoxification of heavy metals. A cDNA encoding type 3 metallothionein (PcMT3) was isolated from the salt stressed leaf cDNA library of Porteresia coarctata (Roxb.) Tateoka (wild rice) that grows well in the heavy metal laden estuarine soils. The PcMT3 cDNA (581 bp) encodes a protein of 64 amino acids. PcMT3 is highly homologous (82 %) to OsMT-I-3a of rice, but is unique from other type 3 plant MTs due to the presence of an additional glycine residue in the C-terminal domain. Analysis of the 5′ upstream region of PcMT3 showed the presence of cis-acting elements like the CG box and STRE previously reported to be involved in gene expression under heavy metal stress. Southern analysis suggested the presence of more than one copy of PcMT3-like sequences in the P. coarctata genome. Analysis of genomic clone of PcMT3 revealed the presence of two introns. A comparison of the genomic sequence of PcMT3 with closely similar type 3 MTs from rice and mangrove species revealed conservation in the number and position of introns. Transcript profiling for PcMT3 in P. coarctata leaves in the presence of Cd, Cu and Zn showed an increase in transcript accumulation.
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