Proc. Indian natn Sci Acad. B70 No.1 pp 1-12 (2004) Plant Based Bioremediation and Mechanisms of Heavy Metal Tolerance of Plants : A Review RK MAITI", JORGE LUIS HERNANDEZ PINERO®, JOSE ANTONIO GONZALEZ OREJA', and DIANA LOPEZ SANTIAGO' (Received on 16 January 2001; Accepted after revision 19 March 2003) Bioremediation of heavy metal-contaminated soils, water and streamsis a great necessity and requires neces sary adaptive measures. Plants with their physical and biochemical mechanisms offer an alternative way of absorption, accumulation and exudation of toxic heavy metals and forming metal-bound compounds. The present paper discusses some basic principles of plant based bioremediation and the biochemical mechanisms of plant resistance to heavy metals. Several hyperaccumulator plant species are being utilized in the bioremediation of heavy metals from soils and water environments. These hyperaccumultors absorb heavy metals, translocate them through tonoplast and accumulate in vacuoles, thereby, protecting cell metabolism from metal toxicity. Some chelate agents acided to contaminated soil promote the uptake of heavy metals by non-hyperaccumulor plants. The mechanisms of heavy metal tolerance may range from exclusion, inclusion and accumulation of heavy metals by plants depending on plant species. The other mechanisms involvesynthe- sisof metal-binding proteins, metallothionin such as Hg, Pb, Cd and phytochelatin such as Cd, Zn, Cd, Pb, and. Hg. Therefore, knowledge on the physiological and biochemical mechanisms of plants for tolerance and accu- :miilation of heavy metals may provide alternate strategies to adopt measures for the purification of soil con- taminated with heavy metals. Biotechnology and genetic engineering has made a significant progress in bioremediation of heavy metals. A number of transporters such members of Zip family have been identified as possibly playing a role in metal exclusion, metal uptake and detoxification. Key Words: Plants, Bioremediation, Heavy metals, Biochemical mechanisms, Tolerance, Phytomining, Introduction A gradual increase of accumulation of heavy metals in the soils of many agricultural fields owing to the discharge of industrial and municipal residual wastes has caused a serious problem to the crop production and hazards to human and animal health. Therefore, remediation of excessive soil metals in situ is receiving great attention, because the alternative, soil removal and replacement is very expensive (Kukier et al. 2000). Bioremediation plays an important role in situ bioremediation of soil contaminated with heavy metals (Bidwell G 1998, Erickson et al. 2000). The problem of soil pollution in USA incurred a cost of 300 billions of dollars in the techniques of remediation (Raskin et al. 1997). The use of plants in remediation, which is usually named Phytoremediation is a cost- effective means to reduce the risk from exposure to hazardous constituents at waste and spill sites (Salt & Rauser 1995, Raskin et al. 1994, Cunningham & Ow 1996). The use of metal- accumulating plants to clean up soil and water contaminated with toxic metals is expanding rapidly in several countries. Certain chelating agents facilitate metal uptake by soil grown plants. The techniques of phytoremediation is being developed as a potential remediation in many of the contaminated sites in the US and other countries (Raskin et al. 1997). This method, though not widely utilized, may be effectively used in industry to treat contaminants at a substantially lower operating cost than more traditional remediation techniques. However, operators must have good knowledge to apply phytoremediation successfully (Beath 2000). Several studies have demonstrated that some plants have the capacity to tolerate high levels of heavy metals without causing any remarkable T. Departamento de Quimica y Biologia, Universidad de las Américas, Santa Catarina Mair, 72820 Choluta, Puebla, México. 2. Postgraduate Division, Biology Faculty, Universidad de Nuevo Le6n, A.P. F-16. 66450 San Nicolis de los Garza, N.L.. México, Corresponding Author: E-mail: rmsiti@mail udlap.mx; Tel: (22) 292608; Fax: (22) 2920962 toxic effects on its metabolic functions. The knowledge on the physiological and biochemical responses may help to adopt different strategies of purification and improvement of the environ- ment through use of plants which tolerate and accumulate high levels of heavy metals. Reviews on bioremediation have been made by Shimp et al. (1993) and Anderson (1993). Raskin and Ensley (2000) have made an excellent review on phytoremediation of toxic metals using plants. The present paper gives a brief review of the biochemical basis of mechanisms for tolerance to the exposure of heavy metals in land plants and recent developments on plant based biore- mediation. Basic Principles of Phytoremediation Phytoremediation may offer a viable solution to the problem of contamination and has drawn, great attractions of environmental scientists. For phytoremediation of metal-contaminated soils, it is essential to understand the interaction between metal-tolerant plant species and soil chemical properties controlling the bioavailability of metals. Phytoremediation is based on the fact that a living plant can be considered as a solar-driven pump, which can extract and concentrate toxic elements from the contaminated soil (Raskin et al. 1997). This involves principally three processes (Brooks et al. 1979, Baker & Brooks 1989), 1) phyoextraction involving extraction of heavy metals from the soil; several wild plants concentrate high amounts of heavy metals in their foliage; 2) phytofiltration (or rhizofiltration) involving of acquatic and semiaquatic plants in various water purification system; 3) phytovolati- zation involving volatization of some toxic metals like Se, As and Hg. Barter (1999) made a review on phytoremediation describing techno-logies used to employ naturally occurring or genetically created plants to remediate a hazardous waste site. In fact, the phytoremedia-tion applies the following basic techniques: rhizofiltration, phytoextraction, phytotrans-formation, phytosti- mulation, and phytostabilization. Species Hyperaccumulators of Heavy Metals Some plant species have capacity to grow in the soill, contaminated with heavy metals and have capacity to accumulate elevated amount of heavy RK Maiti et al. metals (hyperaccumulation) as an ecological adaptation in metaliferous soil (Shaw 1990, Maywald & Weight 1997). Though Brassica sp. Is. not considered as hyperaccumulators, Brassica juncea was identified as a valuable plant for removing heavy metal from soil by Raskin et al. 1997. In a study it was observed that Phaseolus vulgaris accumulated good amount of Pb and Cd (Garay et al. 2000) Plants with capacity to accumulate high concentrations of heavy metals (‘hyperaccumu- ators’) can be used for phytoremediation, for the removal of contaminants from soils. Preliminary results show that Nyssa isa hyperacu-mulator of Co and Ni that can even be used as a feed supplement providing sufficient Co to alleviate Co deficiencies in sheep (Robinson et al. 1999 a, b, c). Willows (Salix species) are able to accumulate high levels of heavy metal ions such as cadmium and zinc. Results have demonstrated the capacity of willow in phytoextraction (Greger & Landberg 1999). In another study, poplar trees sampled in situ contaminated site near the town of Auby, Northern France was found to accumulate significant quanties of Cd. In pot experiments, the clone accumulated more Cd than the poplar clones. Therefore, poplar and willow are hyperaccumulators of Cd, and have been recommended for phytoremediation of Cd contaminated soils (Robinson et al. 2000). Metal hyperaccumulator plants like Thlapsi caerulescens are used for phytoremediation of soils polluted by heavy metals. Robinson et al. (1998) studied the uptake of Cd, Zn, Pb and Mn by Thlapsi caerulescens in plants growing over Pb/Zn mine wastes. They reported that bioaccumulation coefficients were in general higher for Cd than for Zn, and concluded that phytoremediation using Thlapsi caerulesscens ‘would be totally reasonable for low levels of Cd, but not for remediating elevated Zn concentrations within an economic time frame (<10 yr). Thlapsi _goesingense is a hyperacumulator of nickel (Salt 2001). Free hystidine is a metal chelator in plants which helps in the accumulation of Ni (Kramer et al. 1996) Nickel is translocated through cellular membrane and subcellular localization of Ni in the vacuoles is observed in Thlapsi species, thereby leading to hyperacumulation of this metal in plant cell of this species. (Kramer et al. 2000). It has been reported