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Influence of Irrigation with Reclaimed Water on Antioxidant System of Maize and Soybean


Feb. 2010, Volume 4, No.1 (Serial No.26) Journal of Life Sciences, ISSN 1934-7391, USA

Influence of Irrigation with Reclaimed Water on Antioxidant System of Maize and Soybean

Zhanbin Huang1, 2, Wenying Li1, Xiaoqing Wang1, 3, Zhihua Jiao1, Yang Cao1, Shengpeng Jing1 and Zhanxia Miao1
1. College of Chemical and Environmental Engineering, China University of Mining and Technology-Beijing, Beijing 100083, China 2. College of Life Sciences, Henan University, Kaifeng, Henan 475001, China 3. Department of Environmental Engineering and Chemistry, Luoyang Institute of Science and Technology, Luoyang 471023, China Received: October 21, 2009 / Accepted: December 11, 2009 / Published: February 28, 2010. Abstract: Irrigation with reclaimed water is an important way to deal with water shortage of agricultural production and solve the problem of sewage pollution. The response of crop antioxidant enzyme system is an early-warning indicator of environmental factors changes. Compared with raw wastewater and used fresh water as controls, this research studied peroxides (POD), superoxide dismutase (SOD) activities and malondialdehyde (MDA) content in leaves from maize (Zea mays L.) and soybean (Phaseolus vulgaris L.) with pot culture under irrigating with reclaimed water which with secondary and third treatment. The results showed that secondary reclaimed water had some adverse effects on antioxidant system of maize in seedling stage, and the influence decreased in later stage of maize; effect of the third reclaimed water on antioxidant system of maize was not significant. Irrigation with reclaimed water has little influence on antioxidant system of soybean in seedling and reaping stage, but it could bring some oxidative stress in blossom stage. As a result, irrigating maize with second or third reclaimed water is safe to the growth of maize and soybean, but it is suggested that second reclaimed water should not be used in seedling stage of maize and reclaimed water should not be used in the second stage of soybean. Key words: Reclaimed water, maize, soybean, peroxidase (POD), superoxide dismutase (SOD), malondialdehyde (MDA).

1. Introduction
Water is a very limited natural resources. As China’s economic development, problems of water resources shortage, especially agricultural water resources shortage is increasingly serious. At the same time, direct discharge of waste water or reclaimed water, not only waste water resources, but also pollute water bodies. Therefore, the use of reclaimed water is very important to solving the problem of agricultural water resource shortage. The reclaimed water, also called recycle-water, is the product of urbane sewage or domestic sewage with secondary or above secondary treatment. Its quality is up to some water standards and can be reused as impotable water partly. Meanwhile,
Corresponding author: Zhanbin Huang, Ph.D., professor, research fields: physio-ecology of plant, environmental biology. E-mail: zbhuang2003@163.com.

some nutritious elements such as nitrogen, phosphorus and so on in reclaimed water have economic and environmental benefits. Maize is not only a kind of starch food crop with high light efficiency but also a feeding crop and soybean is an important oil crop and food crop with high protein. Both of them account for large proportion in Chinese agricultural production. Therefore, the study of the influence of irrigation with different reclaimed water on maize and soybean’s growth is very important to research practical application of irrigating reclaimed water for them. The process of reclaimed water used for irrigation is equivalent to a deep purification treatment system since when the nutrients and salt in water are absorbed, the water itself is purified gradually as it penetrates soil profile. But once the concentration of pollutants

Influence of Irrigation with Reclaimed Water on Antioxidant System of Maize and Soybean

33

exceeds the self-purification ability of soil, they will have harmful effect on soil property, growth and quality of crop, even on human’s health. There are many researches about the irrigation with reclaimed water, the water quality requirement change with the species of crop and styles of irrigation. Many nations have set some correlated criteria [1-3]. In China, some researchers studied Greenland, grass and golf course under irrigation with reclaimed water, those results showed landscape irrigation with reclaimed water is feasible at large [4-8]. At present the research of influence on crop’s growth and quality is seldom, research of the influence mechanism is fewer. Some detrimental substances in reclaimed water such as salt, heavy metal, organic and so on may stress on crop then induce excessive active oxygen in crop. To avoid the harmful effect of active oxygen, plant has defensive system of antioxidant enzyme protecting itself from damage by impeding the accumulation of active oxygen through maintaining producing and scavenging of active oxygen in dynamic balance. Superoxide dismutase (SOD) and peroxidase (POD) are two key enzymes in antioxidant enzyme system. SOD is universal in the bodies of animal and plant, it’s an enzyme which clearing superoxide anion free radical by converting it into active oxygen. Extent of damage suffered by Plant in stress and plant’s resistivity to stress are frequently related with SOD activity. POD can clearing active oxygen in plant’s body and maintain membrane’s stability, it’s closely related with respiration, photosynthesis and oxidation of auxin. POD is an indicator of metabolism change in a given stage. Membrane lipid peroxidation frequently occurs when plant organ in stress or ageing. Malondialdehyde (MDA), one product of the process, is index of membrane lipid peroxidation to indicate degree of membrane lipid peroxidation and plant’s response to stress. MDA can hold back activities of SOD and POD, then intensify the lipid peroxidation of membrane [9]. Many researches showed the activities of some antioxidant enzyme such as SOD and POD changed

correspondingly when the plant is in the stress of wastewater irrigation or heavy metal [10-12]. In China researches of crop’s antioxidant enzyme largely focus on the influence of a given pollutant on enzyme activity of blade [13-15] rarely refer to comprehensive influence of reclaimed water irrigation. In this research, crop’s response to different stress is studied in terms of hurting mechanism of active oxygen through measuring MDA, SOD and POD in the leaves of maize and soybean. The influence mechanism of reclaimed water on crop’s growth is probed and provided empirical basis for practical application of agricultural irrigation with reclaimed water and setting some standards.

2. Materials and Methods
2.1 Experimental Device The experiment is set with three factors: water quality, growth stage of crop and species of crop. Water includes original wastewater (influent of wastewater treatment plant, Original), reclaimed water with secondary treatment (effluent of secondary sedimentation tank, Second), reclaimed water with third treatment (effluent of sand filter, Third), fresh water (tap water of wastewater treatment plant, Fresh). Four levels are presented in Table 1 and maize and soybean are used as experimental crops. The experiment was replicated more than 3 times. The experiments were conducted at Beijing Gaobeidian wastewater treatment plant with potculture method, Original, Second and Third were got directly from the plant. The maize used in the experiment is China agricultural university 108 and the soybean is Zhonghuang 13. The soil is top soil, procured from experimental establishment of Beijing Hydraulic Research Institute which located in Tongzhou district of Beijing. After air drying, screening soil was put in pots (white, PVC, 16 kg soilpot-1). Before planting the soil was fertilized with urea (account for 0.01 percent) and sterilized by spraying 2,000 mL phoxim solution (40%). Planed six

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Influence of Irrigation with Reclaimed Water on Antioxidant System of Maize and Soybean

Table 1 Water-different quality of irrigation water in the experiment. Chemical composition (mg.L-1) BOD5 CODCr SS TN TP ClPb Cd Zn pH EC/μS.cm-1 Fresh 0.94 2 4 7.81 0.02 45.9 nd nd 0.02 7.77 491 Secondary 9.25 38.1 12.2 30.1 1.95 133 0.31 0.01 0.13 7.85 1199 Third 5.85 23.8 5.55 20.05 2.02 143 nd nd 0.05 7.49 1184 Raw sewage 202.3 411.1 354.3 58.84 6.02 161 0.28 0.02 0.24 7.89 Irrigation water standard (dry farming) 150 300 200 30 10 250 0.1 0.005 2.0 5.5-8.5 1000

seeds of maize or soybean in every pot (two seeds each cave, three caves each pot) and remained three after grown. Weighed the pot and recorded it every other day, sprayed water after the maximum water-holding capacity descended to 50-60 percent and recorded the amount of spraying water. The movable rainout shelters were used to shield the plants from rain in rainy days and must be put away otherwise. The experimental spot was managed in real time and the crop conditions was recorded regularly if encountered by pests, diseases and lodging. 2.2 Contents and Methods of Measuring The maize leaf samples were collected at different stages of seedling, elongation and reaping and the soybean leaf samples collected at stages of seedling, blossom and reaping with each treatment in three replications. Weighed 0.5 g fresh leaf and put it in ultra-low freezer after fixed with liquid nitrogen, then measure the activities of SOD, POD and content of MDA in different leaves, SOD activity was assayed with nitroblue tetrazolium (NBT), POD activity with guaiacol colorimetry and MDA content with thiobarbituric acid colorimetry [15-16]. Data processing were carried out by analysis of variance (ANOVA) with software SPSS. Significant differences among means were determined by LSD at P<0.05.

3. Results and Discussion
3.1 Influence of Reclaimed Water on MDA Content in Maize and Soybean MDA content in leaves of maize were presented in fig. 1. In seedling stage MDA content of plant which irrigated by Second was significantly lower than the plant which irrigated with Original while significantly greater than the plant irrigated with Fresh. No significant difference of MDA content was observed in seedling stage between the plant irrigated with Third and Fresh. In reaping stage, MDA content of plants irrigated with Original and Second were significantly greater than plants irrigated with Third and Fresh. Respect to the whole growing season, MDA content in the plants irrigated with Original and Second first increased then decreased compared with that in the plant irrigated with Fresh. That illustrated irrigation with Original and Second had oxidation stress on maize to varying degrees. To soybean, no significant difference of MDA content was observed between in those plants irrigated with Original, Second, Third and in those plants irrigated with Fresh in seedling and reaping stages. In blossom stage, MDA content in the plants irrigated with Second and Third was markedly lower than in the plant irrigated with Original while greater than in the

Influence of Irrigation with Reclaimed Water on Antioxidant System of Maize and Soybean

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MDA concentration,umol/L

3 2.5 2 1.5 1 0.5 0
maize in seedling stage maize in elongation stage maize in reaping stage soybean in seedling stage soybean in bolssom stage soybean in reaping stage

O S T F

Fig. 1 Water-quality effect on the MDA content of maize and soybean.
450 400 350 300 250 200 150 100 50 0
maize in seedling stage maize in elongation stage maize in soybean in soybean in reaping stage seedling stage bolssom stage soybean in reaping stage

SOD activity,U/gFW

O S T F

Fig. 2 Water-quality effect on the SOD activity of maize and soybean.

plant irrigated with Fresh. That revealed irrigation with Original, Second, Third had little oxidative stress on soybean in the stages of seedling and reaping but had adverse effect on it in blossom stage. This difference may be related to different reaction mechanisms of soybean to harmful ingredients in Original and reclaimed water. 3.2 Influence of Irrigation with Reclaimed Water on SOD Activities in Maize and Soybean In seedling stage, no significant difference of activities of SOD in maize leaf was observed between in the plants irrigated with Second and Third and the plant irrigated with Fresh while both of them were higher than in the plant irrigated with Original. In elongation stage there is no markedly difference among all treatments. In reaping stage, no significant difference of SOD activities was observed between in maize irrigated with Second and Third and in the maize

irrigated with Fresh but SOD activities in the former two were markedly higher than in the maize irrigated with Original (Fig. 2). MDA in maize irrigated with Original in seedling stage increased and produced more active oxygen radicals which enhanced activity of SOD. In latter stage, influence of pollutants in Original on maize reduced so there was no significant difference of SOD activities between in the maize irrigated with Second and Third and in the maize irrigated with Fresh. In seedling stage, SOD activity in soybeans irrigated with Original significantly was higher than in the soybean irrigated with Fresh while no significant difference of SOD activities was observed between in the soybeans irrigated with Second and Third and irrigated with Fresh. In blossom stage, SOD activity in soybean irrigated with Original was markedly lower than in the soybean irrigated with Fresh but no significant difference of SOD activities was observed

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Influence of Irrigation with Reclaimed Water on Antioxidant System of Maize and Soybean

140 120 100 80 60 40 20 0
maize in seedling stage maize in elongation stage maize in soybean in soybean in reaping stage seedling stage bolssom stage soybean in reaping stage

POD activity,U/min*gFW

O S T F

Fig. 3 Water-quality effect on the POD activity of maize and soybean.

between in soybean irrigated with Second and Third and in that planted with Fresh. In blossom stage, the trend of SOD activities was opposite to that of MDA content that’s probably because increase of MAD content inhibits activity of SOD. In reaping stage, no significant difference of SOD activities in soybean was observed among all treatments, and that revealed reclaimed water had little influence on soybean in reaping stage. 3.3 Influence of Irrigation with Reclaimed Water on POD Activities in Maize and Soybean In seedling and reaping stage, there was no significant difference of POD activities between in maize irrigated with Original, Second, Third and in the maize irrigated with Fresh. Only in elongation stage, significant difference of POD activities was observed between in maize irrigated with Original, Second and in the maize irrigated with Fresh while the difference of POD activities between in maize irrigated with Third and irrigated with Fresh was not significant. POD activity in maize in elongation stage is twice as that in seedling stage while in reaping stage that is ten times as that in seedling stage (Fig. 3). In seedling and reaping stages, POD activities in soybean irrigated with Original, Second, Third were not obviously different from that in the soybean irrigated with Fresh. In blossom stage, POD activity in the soybean irrigated with Original was the lowest and it was significantly different with that in the soybean irrigated with Fresh. Comparatively, POD activity in

soybean progressively increased in the sequence of seedling, blossom and reaping stage but the amplitude was smaller than that in maize (Fig. 3). The change was similar to that of SOD activity in soybean that because reclaimed water had little influence on the soybean irrigated with Original and Second in seedling and reaping stage while had great influence on it in blossom stage.

4. Conclusions and Discussions
The research showed that irrigation with Original and Second had great impact on antioxidant system and this influence decreased as maize grew. Moreover Second was good for the maize’s growth in later stage. The detrimental elements in Third to maize’s growth in seedling stage had almost been removed so it had no significant influence on maize’s antioxidant system in seedling stage. Many factors such as ion, microorganism, plankton, heavy metals will have oxidative stress on maize and soybean in seedling stage. MDA content in maize irrigated with Second was greatly lower than that in the maize irrigated with Original. It showed that some elements having oxidative stress on maize had been removed in some degree. But the Second still had adverse influence on maize in seedling stage which displayed by increase of MDA content and decrease of SOD activity in seedling stage. Along the growing process, this influence reduced , especially in reaping stage MDA content in maize irrigated with Second even lower than that in the maize irrigated with Fresh.

Influence of Irrigation with Reclaimed Water on Antioxidant System of Maize and Soybean

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That maybe in later growing stage, maize needs more nutrition and is resistant to external damage with more perfect immune system. No significant difference of MDA content between in maize irrigated with Third and in the maize irrigated with Fresh was observed. MDA content in maize irrigated with Third even tended to increase .It revealed that Third had almost removed unfavorable elements for maize’s growth in seedling stage and even can promote maize’s growth. Irrigation with Second and Third had little influence on soybean’s antioxidant system in seedling and reaping stages but had great effect on it in blossom stage even out of its defensive system’s tolerance .That is indicated by MDA content increased significantly in soybean and SOD activity declined in blossom stage compared with those in soybean irrigated with Fresh .That’s maybe related with soybean’s response mechanism to external stress and subject to further research. Respect to the whole growth process, Third can be used as irrigation water for maize in all stages, Second can be used for maize in the growing stages after elongation stage. To soybean, Second and Third can be used in seedling and reaping stages but should be forbidden in blossom stage.

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Acknowledgments
This paper was supported by the National High Technology Research and Development Program of China (863 Program, No. 2006AA100205) and National Natural Science Foundation of China (No. 50679080).

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