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A B S T R A C T1
SESSION-I
BIOINDICATION and BIOREMEDIATION
A B S T R A C T3
SI/L-1A Review of the Role of Macrophytes in Nu-trient Removal using Constructed Wetlands
Margaret Greenway
School of Environmental Engineering, Griffith University, Nathan,Queensland 4111, AUSTRALIA, Email: m.greenway@griffith.edu.auThere are basically two types of constructed wetlands - Free WaterSurface Systems (FWS) and Subsurface Flow Systems (SSF). FWSwetlands are composed of shallow (20-50 cm) vegetated channelsor basins and deeper (50 cm - 2 m) open-water ponds. SSF wetlands,also known as reed-bed or root-zone wastewater treatment sys-tems, are gravel and/or soil/sand-filled trenches, channels or basinswith no standing water, and support emergent vegetation. FWS sys-tems are more suitable in subtropical/tropical conditions where year-round plant growth occurs. SSF systems are most prevalent temper-ate regions. The effectiveness of water-quality improvement is de-pendent upon an array of complex and interacting processes, whichcan broadly be classified in three categories - physical, biological andchemical. Most processes are facilitated by the wetland vegetation.The wetland plants referred to as macrophytes in constructed wet-land technology, remove soluble inorganic nutrients. However theinteraction between macrophytes and microbes is essential for nitro-gen removal. In aerobic microenvironments around the rhizosphere,nitrification of ammonium occurs. These nitrates can then be taken updirectly by the roots. The dead organic matter of macrophytes pro-vides a carbon source for denitrifying bacteria. The macrophyteseither directly or indirectly play an important role in nutrient removaland storage. The removal of soluble inorganic nitrogen and phospho-rus via absorption from either the water column or the sediment,assimilation and storage in plant tissue is a direct mechanism of nutri-ent sequestration. The provision of plant surfaces (leaves, stems androots) for attached microbiota, epiphytic microflora and associatedbiofilm communities enables microbial assimilation, transformation andstorage of nutrients.
Key words:
Biomass, constructed wetlands, macrophytes, nutri-ents
SI/L-2Detoxification of Heavy Metal Ions and Or-ganic Xenobiotics in Plants
Klaus Lendzian, Andreas Beck, Ralph Blum, Jana Wünschmann and Erwin Grill*
Technical University of Munich, Centre of Scientific ResearchWeihenstephan, Department of Bioscience, Institute of Botany, AmHochanger 4, 85350 Weihenstephan, GERMANY, Email:grill@wzw.tum.dePlants can face a number of foreign (xenobiotic) compounds duringtheir life. These xenobiotics include ions of heavy metals such aslead, cadmium or copper (beyond the physiological relevance), aswell as toxins of microorganisms and agrochemicals. A central role inthe plant responses towards these challenges plays the tripeptideglutathione (GSH) by (i) combating metal ions-induced oxidative reac-tions, (ii) binding heavy-metals via GSH-derived phytochelatins (PCs)which render the toxic ions largely physiologically inactive, and (iii) byGSH-xenobiotic conjugate (GS-conjugate) formation. GS-conjugatesare actively sequestered in the vacuole by the action of ATP-drivenABC-type transporters. While a large body of research has disclosedmetal tolerance mechanisms in metal hyperaccumulators, plants thatcan accumulate heavy metals to levels absolutely toxic to animals(beyond 1% dry weight), that are not mediated by PCs, it is becomingclear that PCs provide a basic form of metal-tolerance and homeosta-sis even in some fungi and the worm
Caenorhabditis elegans
. PCsare generated by the action of a specific dipeptidyl-transferase, PCsynthase (PCS). PCS catalyses the repeated transfer ofglutamylcysteinyl units of GSH onto GSH or PC. PCs bind the heavy-metal ions more avidly than the monothiol GSH. Interestingly, PCS alsocatalyses the turnover of GS-conjugates to glutamylcysteinyl-conju-gates by removing the carboxylterminal glycine. The glutamylcysteinyl-conjugate formation of herbicides has been disclosed in the pioneer-ing work of Gerald L. Lamoureux and identified as being part of acatabolic pathway that is characteristic for plants. Our studies withPCS-knockout lines of
Arabidopsis
unequivocally reveal that PCSconstitutes the major activity for glutamylcysteinyl-conjugate forma-tion. Thus, PCS appear to fulfil a second function in plants, not only inheavy metal detoxification but also in the detoxification pathway ofGS-conjugates.
Key words:
Glutathione, phytochelatin, herbicide, phytochelatin syn-thase
SI/L-3Bioremediation for Heavy Metals throughSymbiosis between Leguminous Plants andRhizobia
Yoshikatsu Murooka
1
*, Akiko Ike
1
, Rutchadaporn Sriprang
2
and Mitsuo Yamashita
1
1
Department of Biotechnology, Graduate School of Engineering, OsakaUniversity, 2-1 Yamada-oka, Suita-shi, Osaka 565-0871, JAPAN, Email:murooka@bio.eng.osaka-u.ac.jp;
2
BIOTEC Center for Genetic Engi-neering and Biotechnology, National Science and Technology Devel-opment Agency, 113 Phaholyothin Rd., Klong 1, Klong Luang,Pathumthani, 12120, THAILANDAccumulation of heavy metals, such as As, Cd, Cr, Cu, Ni, Pb and Zn,is harmful to living organisms in contact with the soil and groundwater.Cadmium contamination in rice grains is one of the important issues inAsian countries. A novel bioremediation system, “symbiotic engineer-ing”, using symbiosis between leguminous plants and rhizobia wasdeveloped.We synthesized the tetrameric metallothionein gene (
MTL4
, Murooka
et al.
, Biocatalysis Biotransformation, 19: 399-412, 2001) and cDNAencoding the phytochelatin synthase gene (
AtPCS
) from
Arabidopsis
.The
MTL4
and
AtPCS
genes were fused to the
nifH
promoter, whichgenerated nodule-specific expression of these genes in
Mesorhizobium
huakuii
subsp.
rengei
B3, which can infect and formnodules on Chinese milk vetch,
Astragalus sinicus
(Sriprang et al.
Appl. Environ. Microbiol
69: 1791-1796).By the expression of the
AtPCS
gene, phytochelatin [(g-Glu-Cys)
n
-Gly ; PCs] was formed in
M. huakuii
subsp.
rengei
strain B3 andincreased the ability of cells to bind Cd
2+
by 9- to 19-fold approximately.When recombinant strain B3 established the symbiotic relationshipwith
Astragalus sinicus
, the symbionts increased Cd
2+
accumulationin root nodules by 1.5-fold. The expression of the both
MTL4
and
AtPCS
genes showed additive effect on cadmium accumulation innodules. Furthermore, we expressed the
AtPCS
gene and iron-regu-lated transporter (IRT1) in strain
B3 to stimulate the uptake of cadmiuminto the nodules. The expression of both the
AtPCS
and
IRT1
genes inrecombinant strain
B3 increased the ability of cells to bind Cd
2+
1.7 to2.5-fold compared to cells expressing only
AtPCS
. In rice paddy soil,addition of recombinant strain B3 significantly increased the accumu-lation of cadmium in roots and nodules of
A. sinicus
.Thus, this system uses the advantages of both plants and rhizobia, inparticular, engineered genes can be transformed to plants throughinfection with recombinant bacteria.
Key words:
Environment, Heavy metals, Cadmium, Rice, Symbiosis,
Astragalus sinicus
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