0278-6915(95)00004-6 Fa Chem. Tosie. Vol. 3b, No. $, pp. 398-397, 1998 Copyright © 1995 Elsevier Science Lid Printed in Great Britain. All ihts reserved (0278-6915)98 $9.50 + 0.00 Studies on Uptake of Trivalent and Hexavalent Chromium by Maize (Zea mays) S. MISHRA*, V. SINGH*, S. SRIVASTAVA*, R. SRIVASTAVAS, M. M. SRIVASTAVA‘, S. DASS*, G. P. SATSANGIt and S. PRAKASH} “Department of Chemistry and {Department of Botany, Dayalbagh Educational Institute, Dayalbagh, ‘Agra-282 005, India (Accepted 20 December 1994) Abstract—Pot culture experiments were carried out to study the uptake and translocation of chromium from irrigation water when supplied in its trivalent and hexavalent states to maize plants grown in soil and sand culture, The uptake of chromium was observed to increase with increase in the concentration for both oxidation states of chromium. For the root, the observed order of uptake was CrHlI sand > CrVI sand > CrlII soil > CrVI soil, whereas in the lower shoot, upper shoot and fruit the order was CrVi sand > CrV1 soil > CrIf1 sand > Crill soit. Significantly high uptake of chromium by roots in the send culture (Crlll treatment) is attributed to the effect of root exudates and degradation products on the mobilization of Crill. In aerial parts of the plants a higher uptake was observed when the initial supply was CrV1, The trends observed are explained on the basis of the redox behaviour of chromium. INTRODUCTION The behaviour of chromium (Cr) in water and soil and its subsequent bioavailability have been the subject of active investigation because of the existence of chromium in two environmentally important oxi- dation states and the ease with which it can complex with naturally occurring chemical species in the soil Studies on various aspects of plant uptake of chro- mium have been reported by several workers (Bartlett, 1991; Bartlett and James, 1988; Cary, 1982; EPA, 1984; NRC, 1974). tis believed that at natural pH, CrV1, being water soluble and of smaller than the hydrated Crifl ion, readily penetrates cell walls and exhibits toxic be- haviour. CrIll, on the other hand, has a strong tendency to become hydrolysed and adsorbed, finally becoming immobilized. The hydrated CrIl¥ cation does not pass through the cell membrane, even at low pH (Cary et al, 19772,b; Lahouti and Peterson, 1979). Crlll, however, when organically bound at biological pH values, is known to penetrate the cell wall (Bartlett and James, 1988). The reduction of CrVI to rill by organic matter in soil has been reported by Bartlett and Kimble (1976) and the oxidation of CrIII has been reported by Bartlett and James (1979). Studies on the plant uptake of chro- ‘mium have shown that uptake and translocation by plants is, in general, low, with maximum retention in {Author for correspondence, the roots (Turner and Rust, 1971; Peterson and Griling, 1981), The uptake of chromium by plants has been stud- ied extensively in hydroponic systems as well as when chromium has been supplied as a soil amendment. However, the plant uptake of chromium, when it is fed through irrigation water, has not yet been studied. Recently, we have reported results of a study on the uptake of Crll] and CrVI through irrigation water by the onion plant, grown in soil and sand culture (Srivastava et al., 1994). The present paper describes a study on the uptake of chromium by the maize plant (Zea mays). As quartz sand is an inert medium, ‘a comparison between the results of soil and sand culture experiments is likely to highlight the role of soil, which can act as an oxidizing or reducing, as well as a complexing, medium in the uptake of chromium by plants (Bartlett and James, 1988). Sand culture experiments may also shed light on the possible role of root exudates and root degradation products in modifying the chemistry of chromium, particularly in the mobilization of Crill. MATERIALS AND METHODS Pot culture experiments were performed under laboratory conditions using plastic containers with 2.5 kg soil and sand separately. A basal dose of NPK ‘was initially applied to the soil. About cight seeds were sown in each pot and after 10 days from germination four plants per pot were retained for studies, Similar experiments were carried out in sand394 ‘Table |. Plant tissue concentration of chromium in various parts of maize plants treated with CrITT and CrVT, grown in soil and sand culture Chromium concentration Tm plant issue sie dry weight) __ In irrigation cen crv Plant water ————— _part_(ugiml)_Sand_Soil_Sand_Soil_ g 05 » 0 @ 1s 10 mr 1036 50 250 aS 250 3y75 12761580 Ls os 2 3 2 10 s 9 2 2 50 m% 3% 12s 250 me hae RS us 05 1 29 6 10 : 1 6 6 30 7 4h 6 50 6% 2 60 tao G os 1 2 44 1 4 5 7 7 50 3s 7 1% 250 i a Rovwot US lower shoot US=upper shoot G= eran (high-quality silica quartz), feeding with complete ‘nutrient solution and iron citrate (Hoagland et al., 1938), Both sand and soil pots were irrigated when necessary and the pH values of soil and sand in the pots were close to 7.8 and 7.2, respectively. Plants were allowed to grow for about 70 days and at the time of maturity (when the maize grain reached the “milky-ripe’ stage) the pots were transferred to fume- hoods for feeding with *!Cr-tabelled Crlll and CrVI itrigation water. It would have been desirable to study uptake at more than one stage of plant growth; however, as the major aim was to study the chro- mium reaching the edible part of the plant, the feeding was done at the milk stage only. “Table 2 Percentage of total intake of CHIT and CVI accumulated in different plant parts ia sand and soil culture Percentage accumulation of tal intake of Cr Cel vi Plant Cr concn in plant part” __Getig.dry mitier) Sand Soil Sand Soil R 05 7s4 69.3 25269 Ls 05 "3 28 473 453 us 05 Wi 2 BL 6 G os 220028 7243 . 0s 309 78174382 R 10 7 138 314 28 Ls to BS 218 7 389 us to 20 48 1S 100 o 10 39 09 4320 + Lo 2 17 485 953 R 50 9 633 390 258 Ls 50 259 Wl 4 S84 us 30 350024 «49173 G 30 38 43 6826 * 50 3 95 400 660 R 250 @s @s m9 MS Ls 250 Ma BI 9 356 us 250 3755 9 4 c 250 90 20 7334 . 280 2104834603 Asin Table | ‘Total percentage of Cr (based on feed value) by the whole plant SS. Mishra er al 3500 (ay 2500 2000 1300 1000 f- 500 600 300 400 300 200 100 300 250 200 Chromium uptake (g/g dry weight) 150 100 so 600 300 400 300 200 100 rr Chromium administered (ug/ml) Fig. 1. Increase in percentage uptake of chromium from irrigation water with increase in the concentration of chro- rium administered (™, Crlll; sand culture; +, Crlll, soil culture; *, CrVI, sand culture; (2, CrVI, soil culture), by various parts of Zea mays plants (a, root; b, lower shoot; ¢, ‘upper shoot; d, grain). Four pots each, of sand and soil, were irrigated ‘once only with 100 ml solution of Crill and CrVI (0.5-25 g/ml), labelled with a known amount of SiCr-activity (a few Ci/pot). The concentrations selected were the upper limits of concentration of chromium that can be found in irrigation water andUptake of Criff and CrlV by maize Uptake and translocation 395 ROOT SYSTEM Uptake and immobilization Oxidized Organic material Ma in root exudates & fresh & root degradation moist soil products & aeration > variations in the uptake of CrIII and CrVI within and around the plant root Labile Crit som i SYSTEM r i i 1 1 1 1 ' hydrolyzed & immobilized system. soils. Chromium chloride and potassium dichromate (AR. grade) were used as salts of trivalent and hexavalent chromium, respectively. A stock solution of Crlll was calibrated by oxidizing it to CrVE and estimating it spectrophotometrically. The required inactive chromium, in their respective oxidation states, suitably oxidized or reduced was mixed with 5\Cr before feeding to plants. ®'Cr was obtained from the Board of Radiation and Isotope Technology (BRIT) BARC, Bombay, India. Both radioactive decay and gamma ray spectra of stock Cr confirmed the purity of the radioisotope obtained. Blank values for chromium determined using atomic absorption spectrometry on soil, sand Jeachates and irrigation water were found to be below the detection limit (approx. 40 ng/ml), indicating that the contribution of chromium from these sources and from *'Cr activity (estimated to be- less than Sng/uCi) was negligible. Illumination of about 600 W/m? was pro- vided by fluorescent tubes in the fumehoods, together with a slow ventilation system. Plants were kept in the fumehood system for 7 days Gaily illumination of 14hr) for chromium uptake. They were then harvested and separated into root (R), lower shoot (LS), upper shoot (US) and fruit (F) and were washed thoroughly with running water, then rinsed with 0.01 HNO. Plant parts were washed five times with water and the pH of the washings tested to ensure that no detectable acidity remained. In order to check external contamination of plant parts, acid washings and final washings of representative samples were assayed for ‘'Cr: there was no detectable contamination. Plant parts were dried in an oven at 60°C for about 48 hr and were cut into smaller parts and stored in labelled plastic packets. Accurately weighed amounts of dried ma- terial (practically all the dried material) in the stan- dard geometry were counted over a planar Nal (TI) detector coupled to a 4K MCA (Canberra Acuspec Card with PC-AT 386). The counting geometry was precalibrated for efficiency with known amounts of 51Cr activity. From the 320 KeV photopeak area, the activity of *'Cr was calculated and expressed as the total amount of chromium in different plant parts/g dry weight. All the activity was corrected for its decay to arrive at an activity at a common time and date. Samples were counted for varying durations (10-120 min) to accumulate at least 2000~3000 counts under the photopeak area, to keep the statisti- cal error in counting below a few per cent. The data represent the means of four plants per concentration for both sand and soil culture with CrlI and CrVI as feed. Correlation coefficients were calculated and the chi-square test applied where appropriate. The calculated radioactivity values represent an average of four plants per concentration of irrigation for both sand and soil culture. It should also be remembered that the final results mentioned in this work refer to chromium per g (total chromium) of dry matter in lower shoot > grain > upper shoot [1]