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Drought stress decreased leaf water absorption capacity and real water content and increase relative water content in genotypes of chickpea. It decreased chlorophyll a and b content. Drought increased peroxidase superoxide dismutase, glutathione reductase, ascorbate peroxidase and catalase in stress groups. Proline content increased drastically index stress condition. Introduction Chickpea is the third pulse crop followed by bean and pea in world production. It is clear that the sowing area, production and yield of chickpea decreased during the last ten years (Ceyhan et al. 2007, Ceyhan et al. 2012a). The main cause of reducing of yield is biotic (Ascochyta rabiei) and abiotic (high temperature and drought) factors. A closed basin Central Anatolian region is one of the most affected regions by drought (Ceyhan et al. 2012a). This region has almost 50% of the total chickpea production in Turkey. Drought considerably affect chickpea cultivation, drought tolerant line of chickpea is none (Singh 1997). Drought is known as the most important abiotic stress factor over the world (Ceyhan et al. 2012a) and it causes loss in yield of the cold climate pulses (Singh 1997). Drought stress in plants induces physiological, biochemical and molecular changes for adaptation (Arora et al. 2002, Kalefetoğlu 2006). Number of work related to drought stress in chickpea is very few. Effect of drought on physiological and biochemical changes chickpea genotypes is reported. Materials and Methods A total of ten genotypes were collected from six local population (Kadinhani LP, Altinekin LP, Hadim LP, Cumra LP, KarapinarLP and Beysehir LP) from city of Konya, two standard cultivar (Canitez and Kusmen-99) and two genotypes (22117 and 22223) from ICARDA (drought tolerant). The pots which had a volume of 1 liter (14 × 13 cm) were washed and sterilized for planting in greenhouse. The seeds of genotypes were exposed to 5% sodium hypochlorite for 3 min and then washed for 3 times with de-ionized water for sterilization. Subsequently, seeds were sown in uncontaminated pots. The experiment was conducted in “Randomized Plots Factorial Design with Two Factors” with three replications in a fully controlled research greenhouse in Department of Soil Sciences and Plant Nutrition, Faculty of Agriculture, University of Selcuk in 2009. Sowing was made by hand on 15th of February 2009. The top sides of pots were closed for one week after sowing and placed in the greenhouse at 25C temperature and 40 50% relative humidity. The top sides of pots were opened after seeds were germinated. The seedlings were grown in the greenhouse under 25C and 40 50% relative *Author for correspondence: . 178 GÖKMEN AND CEYHAN humidity conditions for 40 days after emergence. Then, the plants were classified as control (0 Day), and three stress groups (3rd, 5th and 7th day) (Ceyhan et al. 2012 b). Drought stress was applied by non-irrigation for 3, 5 and 7 days. The harvest was made in the same order with stress groups which was started with 0 day (40 days after emergence), and following 3rd, 5th and 7th days of stress (Ceyhan et al. 2012 b). Some physiological and biochemical analysis were made on the leaf tissue of harvested stress and control groups in trial. Leaf water absorption capacity (LWAC), relative water content (RWC) and real water content (REWC) were determined according to the method of Clarke and McGaig (1982) and Farrant (2000). The amount (mg/l) of chlorophyll a, chlorophyll b and total chlorophyll (a+b) were determined according to Lichtenthaler (1987). Leaf samples (0.5 g) were frozen (separately) in liquid nitrogen and stored in deep freezer (80C). Likewise, 0.1 g leaf samples were also frozen in liquid nitrogen and stored in deep freezer (80C). 0.5 g leaf samples were homogenized in liquid nitrogen with %2 w/v polyvinylpolyprrolidone (PVPP) and 1 mM EDTA, pH 7, 8 and 50 mM Na-phosphate buffer medium. After filtration, centrifuge was made on + 4°C, 14 000 rpm for 30 min. These processes were made separately for each of the peroxidase (POD), superoxide dismutase (SOD), glutathione reductase (GR), ascorbate peroxidase (APX), catalase (CAT) and proline analysis (Kumar and Kahn 1982, Beauchamp and Fridovich 1971, Bates et al.1973, Foyer and Halliwell 1976, Nakano and Asada 1981, Bergmeyer 1970). Results and Discussion LWAC of stress groups showed that the genotypes Karapinar LP (0.0063 g/gh), Kusmen 99 (0.0060 g/gh) and 22223 (0.0054 g/gh) had more LWAC. Means of genotypes showed that the highest LWAC belongs to control group (0.0132 g/gh). As increasing in drought stress; LWAC decreased (Table 1). An important level of reduction in the LWAC of the genotypes occurred by application of drought stress compared with their own controls. The same result was also reported by Ceyhan et al. (2012b). RWC of stress groups showed that the highest value was observed in the genotype Canitez (81.89%) while the lowest value was observed in genotype Beysehir LP (66.40%) (Table 2). The RWC values decreased compared to their control genotypes Kadinhani LP, Altinekin LP, Hadim LP, Karapinar LP and Beysehir LP but that decreased in the other genotypes (Table 2). Many researchers implicated that drought had negative effect on RWC in the plants (Anyia and Herzog, 2004, Ceyhan et al. 2012b). RWC decreased in bean even after closing of stoma (Costa Franca et al. 2000) and in chickpea (Kalefetoğlu 2006) following stress application. On the other hand, Turkan et al. (2005) described the high RWC in the drought-resistant Phaseolus acutifolius was due to high content of proline. REWC of stress groups showed that the highest value was observed on the genotype Kusmen 99 (83.41%). Means of the genotypes showed that the highest REWC value (84.47%) was found on control group (Table 1). The REWC values decreased comparing with their control of the genotypes Kadinhani LP, Altinekin LP, Hadim LP, Karapinar LP and Beysehir LP while an increasing was observed in the other genotypes (Table 2). Kalefetoğlu (2006) working with chickpea observed reducing in REWC compared to controls by increasing the drought. Effect of drought on chlorophyll content: Chlorophyll a of the stress groups ranged from 1.62 (Kadinhani LP) to 3.78 mg/l (Kusmen 99). Means of the genotypes showed the highest value (2.640 mg/l ) in control group while the lowest value (2.08 mg/l) was observed on 7 days stress group (Table 1). All the genotypes showed an important decrease for chlorophyll a content as from 3 days compared with their own control group (Table 1). EFFECTS OF DROUGHT STRESS ON GROWTH PARAMETERS 179 Chlorophyll b of the stress groups produced the highest value Canitez and it was followed by Kadinhani and Altinekin (2.518, 1.960 and 1.935 mg/l, respectively). The lowest value occurred on Karapinar LP with 0.842 mg/l of chlorophyll b content (Table1). Table 1. Effects of drought on LWAC, RWC, REWC and chlorophyll a, b, a+b inchickpea genotypes. LWAC RWC REWC Chlorophyll a Chlorophyll b Chlorophyll a+b g/gh % % mg/l mg/l mg/l Stress groups Control 0.0132 a* 73.92 84.47 a 2.640 a 1.613 a 4.243 a 3rd day 0.0021 b 76.31 80.69 b 2.520 b 1.464 b 3.975 b 5th " 0.0018 b 78.18 80.69 b 2.380 c 1.301 d 3.681 c 7th " 0.0018 b 76.31 76.30 c 2.076 d 1.386 c 3.463 d

Effects of drought stress on growth parameters, enzyme activities and proline content in chickpea genotypes