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Butachlor Induces Some Physiological and Biochemical Changes in a Rice

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Butachlor Induces Some Physiological and Biochemical Changes in a Rice Pesticide Biochemistry and Physiology 105 (2013) 224–230 Contents lists available at SciVerse ScienceDirect Pesticide Biochemistry and Physiology journal homepage: www.elsevier.com/locate/pest Butachlor induces some physiological and biochemical changes in a rice field biofertilizer cyanobacterium ⇑ Hongzhi He a, Yongjun Li b, Tianfeng Chen c, , Xiaolong Huang a, Qiu Guo a, Shufeng Li a, ⇑ Tianhong Yu a, Huashou Li a, a Key Laboratory of Agro-Environment in the Tropics, Ministry of Agriculture of the People’s Republic of China, South China Agricultural University, Guangzhou 510642, China b Zhongshan Quality Supervision & Inspection Institute of Agricultural Products, Zhongshan 528403, China c Department of Chemistry, Jinan University, Guangzhou 510632, China article info abstract Article history: Butachlor has been widely applied in rice field in China. However, concerns are also raised about its Received 18 September 2012 potential adverse impacts on non-target organisms. In the present study, butachlor was found be able Accepted 25 February 2013 to induce toxic effects on a rice field biofertilizer cyanobacterium Nostoc sp. When treated with 80 mg LÀ1 Available online 7 March 2013 butachlor, significant decline in the growth rate, concentrations of chlorophyll a (Chla), carotenoids (Cars), phycobiliproteins (PBPs) and; the minimal fluorescence yield (F0), fluorescence intensity at the Keywords: J-step of OJIP (Fj), the maximum fluorescence yield (Fm), the potential quantum yield (Qy), the quantum Nostoc sp. yield of electron transport (UE ), the maximum quantum yield of primary photochemistry (UP ), and the Butachlor 0 0 performance index on absorption basis (PI ) (14.2%, 39.5%, 55.5%, 34.8%, 38.5%, 19.8%, 18.7%, 20.4%, Toxicity ABS Antioxidant system 10.1%, 10.3%, and 26.4%, respectively) was observed in Nostoc sp. In contrast, significant increase in Fluorescence Cars/Chla, PBPs/Chla, the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and, glutathione reductase (GR), content of malonaldehyde (MDA), the absorption flux per reaction center (ABS/RC) and the effective dissipation per reaction center (DI0/RC) for 0.45, 0.65, 2.36, 2.47, 1.08, 1.16, 0.87, 0.122 and 0.205 fold was also detected by comparing with the control group. These results demon- strated that high concentration of butachlor could inhibit the growth, synthesis of pigments, and photo- system II (PSII) activities of Nostoc sp., and trigger dramatic intracellular antioxidant response in the cells. Taken together, this study may provide important information on the understanding of the changes induced by butachlor stress in nitrogen-fixing cyanobacteria and the adaptive strategy of the alga. Ó 2013 Elsevier Inc. All rights reserved. 1. Introduction acted by inhibiting the elongase responsible for the elongation of very long-chain fatty acids and geranylgeranyl pyrophosphate cyc- Chloroacetanilide herbicides (e.g. acetochlor, metolachlor and lisation enzymes [3]. As a consequence of the use in large quantity, butachlor) are important chemicals used in agriculture all over contamination of butachlor is now very common in groundwater the world. They were commonly used to control annual grasses and surface-water in China [4], and result in disruption of the envi- and certain broad-leaved weeds in both seeded and transplanted ronment and ecosystems [5]. rice [1]. In China, N-(butoxymethyl)-2-chloro-N-(2,6-diethylphe- Nitrogen is one of the most important factors that limit the rice nyl) acetamide (butachlor) is now one of the three most widely production. Nowadays, chemical N fertilizers are widely applied in used herbicides, with an annual yield of 1 Â 104 t [2]. Butachlor rice field to promote rice yield in China [6]. However, because of Abbreviations: Chla, chlorophyll a; Cars, carotenoids; OD, optical density; DW, dry weight; PBPs, phycobiliproteins; PC, phycocyanin; APC, allophycocyanin; PE, phycoerythrin; SOD, superoxide dismutase; CAT, catalase; POD, peroxidase; GR, glutathione reductase; MDA, malonaldehyde; ANOVA, analysis of variance; DCMU, 3-(3,4- dichlorophenyl)- 1,1-dimethylurea; 2,4-D, 2,4-dichlorophynoxyacetic acid; ROS, reactive oxygen species; GSH, glutathione; PSII, photosystem II; Ft, the instantaneous Chla fluorescence; Fj, Fluorescence intensity at the J-step of OJIP; Qy, the potential quantum yield; M0, approximated initial slope of the fluorescence transient; VJ, relative variable fluorescence at the J-step; UE0, the quantum yield of electron transport; UP0, the maximum quantum yield of primary photochemistry; W0, the probability that a trapped À exciton moves an electron further than QA ; ABS/RC, the absorption flux per reaction center; TR0/RC, the trapping flux per reaction center; ET0/RC, the electron transport flux per reaction center; DI0/RC, the effective dissipation per reaction center; PIABS, the performance index on absorption basis. ⇑ Corresponding authors. Address: Department of Chemistry, Jinan University, Guangzhou 510632, China, Fax: +86-020-85220223 (T. Chen), College of Agriculture, South China Agricultural University, Guangzhou 510642, China, Fax: +86-020-85280211 (H. Li). E-mail addresses: [email protected] (T. Chen), [email protected] (H. Li). 0048-3575/$ - see front matter Ó 2013 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.pestbp.2013.02.009 H. He et al. / Pesticide Biochemistry and Physiology 105 (2013) 224–230 225 the large requirement of fossil-fuel energy for the production of 2.3. Dry weight (DW), growth curves and specific growth rate these fertilizers, the prices of them become higher and higher. At measurement the same time, chemical N fertilizers could post many adverse ef- fects on the environment. Therefore, it is very important to survey For DW determination, 15 mL algal culture were filtered and develop alternate low-cost, assimilable nitrogen sources [7]. through a pre-dried Whatman GF/C paper, washed three times The natural nitrogen resources exhibit unique properties of biolog- with culture media, and the filters containing cells were dried at ical nitrogen fixation for agricultural use. Nitrogen-fixing cyano- 80 °C until constant weight and then weighted. A significant posi- bacteria are abundantly present in the rice field of south China tive relationship between DW and the optical density at 680 nm and are important microbes in maintenance of rice field fertility (OD680) detected by a Shimadzu UV-250IPC spectrophotometer À1 through carbon and nitrogen fixation [8]. Nowadays, microalgal showed [DW (mg L )=À1.94 + 231 ⁄ OD680, r = 0.9998, biofertilizers have been proved to be a useful tool for increasing p < 0.0001] according to the results of preliminary experiment. the productivity of rice production and improving physical and Therefore, the growth was measured at 680 nm at 48-h interval chemical properties of the soil in agricultural system [9]. However, using Shimadzu UV-250IPC spectrophotometer for 12 d. During the large scale use of herbicides in modern rice agriculture has af- the exponential growth phase, the specific growth rate (l,dÀ1) fected cyanobacteria adversely in various ways [10]. could be calculated using the equation: Till now, many studies have been conducted to assess the tox- 1 N icity of butachlor on the growth, photosynthesis and physiology l ¼ ln j ð1Þ j i N of nitrogen-fixing cyanobacteria [11–17]. However, little attention À i À1 has been paid to the antioxidant system and Chla fluorescence where Ni and Nj represented DW (mg L ) at day i and day j, changes in nitrogen-fixing cyanobacteria under butachlor stress respectively. conditions. Therefore, the aim of this study was to investigate the toxic effects of butachlor on the growth, PSII activities, bio- 2.4. Determination of Chla and Cars chemical composition and antioxidant system of a nitrogen-fixing cyanobacterium Nostoc sp. FACHB-85 isolated from paddy fields in Twenty mL algal culture were filtered through 0.45 lm filters South China. and the filter membrane was transported to a 10 mL plastic tube and extracted with 6 mL ice-chilled 80% acetone for 24 h in dark at 2. Materials and methods 4 °C. The solution was then centrifuged at 5000 rpm for 10 min to re- move the cell debris, and the supernatant was collected for determi- 2.1. Chemical, organism and instrument nation of Chla and Cars according to the formulas of Lichtenthaler and Wellburn [19]. The formulas were showed below: The tested herbicide N-(butoxymethyl)-2-chloro-N-(2,6-dieth- CChla ¼ 12:21 Â OD663 À 2:81 Â OD646 ð2Þ ylphenyl) acetamide (butachlor, 99.7%) was purchased from Shanghai Pesticides Research Institute (China). Heterocystous cya- CChlb ¼ 20:13 Â OD646 À 5:03 Â OD663 ð3Þ nobacterium Nostoc sp. (named as FACHB-85) was originally iso- lated from rice field in Hubei Province and obtained from the 1000 Â OD À 3:27 Â C À 104 Â C C ¼ 470 Chla Chlb ð4Þ Freshwater Algae Culture Collection of the Institute of Hydrobiol- Cars 229 ogy, the Chinese Academy of Sciences. Stock culture of the alga Where OD ,OD , and OD represented absorbance at 470, 464, was maintained at 27 °C in sterilized N-free mineral medium 470 646 663 and 663 nm, respectively. BG11 [18] at pH 7.5 with 3000 lx light intensity provided by cool- white fluorescent lamps and a 16 h/8 h light/dark cycle. A Shima- dzu UV-250IPC spectrophotometer was employed for absorbance 2.5. Determination of PBPs determination. For determination of PBPs, samples obtained by filtering 20 mL algal culture were repeatedly frozen at À40 °C and thawed at 4 °C 2.2. Butachlor treatment for 6 times in 0.05 mol LÀ1 phosphate buffer (mixing equal vol- À1 À1 umes of 0.1 mol L KH2PO4 and 0.1 mol L K2HPO4 solutions, Stock solution of butachlor was prepared with analytical grade pH 7.1). And then the solution was ultrasounded for 120 s on a acetone. Firstly, preliminary experiments were conducted to exam- sonicator (YJ92-II, Yong-Jie Experimental Laboratory Apparatus ine the effects of the solvent acetone alone on the cyanobacterium Corporation, Ning Bo, China) equipped with a probe operating at and the results showed that 0.6% (v/v) acetone had no significant 80% in 5-s pulses) in ice-water bath for 10 min.
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