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Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits of Lentils (Lens culinaris L.) Under Hydroponics Conditions



M. Salehpour, A. Ebadi, M. Izadi and Sh. Jamaati-e-Somarin
 
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ABSTRACT

In order to investigate the response of Lentil cultivar Gachsaran to nitrogen and water stress under hydroponics conditions, a factorial experiment based on randomized complete block design was conducted with three replications using Hoagland nutrient solution without nitrogen, at the growth chamber in 2007, Ardebil, Iran. Factors were four water stress levels (0, -0.6, -0.9 and -1.2 MPa) prepared with polyethylene glycol (PEG) and three nitrogen levels (0, 0.5 and 1.0 Mmol). Results showed that all traits were decreased with increasing stress. Both treatments had significant effects on Relative Water Content (RWC), Leaf Chlorophyll Content (LCC), Leaf Area (LA), length, height and dry weight of stem and Total Biomass (TB) and their interaction effects on LCC (before and after stress), LA (p<0.05) and on length and dry weight of stem (p<0.01). The most values of RWC, LCC, LA, length and dry weight of stem and total biomass were obtained using 0.5 Mmol nitrogen is sufficient and higher values is not recommended because of its preventive effect on nitrogen fixation in plant.

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M. Salehpour, A. Ebadi, M. Izadi and Sh. Jamaati-e-Somarin, 2009. Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits of Lentils (Lens culinaris L.) Under Hydroponics Conditions. Research Journal of Environmental Sciences, 3: 103-109.

DOI: 10.3923/rjes.2009.103.109

URL: https://scialert.net/abstract/?doi=rjes.2009.103.109
 

INTRODUCTION

Seed legumes are important sources of vegetable proteins for human and animals and are main crops in agriculture (Dita et al., 2006) and because of their ability to nitrogen fixation (Howieson and Ballard, 2004). Lentils (Lens culinaris L.) is a member of legumes family and does nitrogen fixation in presence of Rhizobium leguminosarum (Schmidtke et al., 2004). Because of high content of protein in legumes, demand for nitrogen is higher than other nutrient elements. So, its deficiency is very critical for plant, because in the lack of this element, chloroplast, chlorophyll and many of metabolic enzymes is disordered. There are two nitrogen sources for legumes including mineral and fixed nitrogen which have been shown to be fixed in Lentil of about 78% by 15N method (Maskey et al., 2001). Moreover, water stress is one of the most important and prevalent environmental stresses that limits farming (Rosales-Sernaa et al., 2004). Drought stress decreases number, size, area and chlorophyll content of the leaf, height and biomass of the plant which could be attributed to vegetative growth decrease such as low production of new leaves, high leaf senescence, decrease in mean leaf size, decrease in plant height and etc. (Pagter et al., 2005). Also, drought stress has adverse effects on nodulation and nitrogen fixation (Athar and Johnson, 1996). The highest fixation is done while soil nitrogen content is low and soil humidity and temperature is favorite. It had been found that drought stress resulted in low nodulation and nitrogen fixation in bean up to 25-30% than control (Verdoy et al., 2004). Based on the time of nitrogen application and root zone conditions, nitrogen can cause early maturing or idiomatically, green-drying of plant under drought circumstances and consequently, decrease of yield. The reason is that water consumption by plant becomes faster and stored carbohydrates, decreases (Sadras, 2002). Perhaps, for this reason the, most farmers stop or decrease nitrogen utilization in drought circumstances. Yang et al. (2000, 2001) reports that increase of nitrogen led to yield increment. Usually in case of desired nitrogen fixation, there is no require to nitrogen application. However, symbiosis between seed legumes and rhizobium have been improved to the large extent, but is sensitive to environmental conditions and stresses, it will extinct. In this condition, nitrogen applications can improve plant resistance and yield against stresses. Antolin et al. (1995) and Solan et al. (1990) have reported same results in alfalfa and soybean and stated that plants which received nitrate, were more tolerant to dehydration against nitrogen fixator plants. This improvement in plant growth relates to more produced aerial parts such as crown, stem and leaf area and more nitrogen fixation in roots. Generally, environmental factors such as temperature call not be controlled by farmers but nutrient deficit and weak nitrogen fixation can be improved by infection, fertilization irrigation and other management methods (Lindmann and Glovir, 2003). Many researchers showed that legumes which fed by nitrate or ammonium, had better growth than those relying on nitrogen fixation (Silsbury et al., 1986; Sprent and Thomas, 1984). This is because of effect of nitrogen on lengthening the stem, increase of number and area of the leaves increase of LA which may arising from increase of number or area of the leaves caused by nitrogen utilization. Response of plant to drought stress pertain to water deficit and comprises of shot-term and long-term reactions. RWC, LCC, MSI, TB and plant vegetative growth such as number and area of the leaf, plant height and etc are used ass criterions representing plant tolerance to drought stress. Costa-Fraca et al. (2000) reported that RWC in bean leaves was reduced under water deficit. High values of leaf RWC in tolerant cultivars may attributable to lower water losing from stomata and/or higher water absorption by roots as a result of special mechanism exerted by plants (Jiyang and Hung, 2001). LCC in one of key factors determining photosynthesis rate and dry matter production, as well (Ghosh et al., 2004). Also, MSI is a criterion representing plant resistance against water deficit and its value decreases in this condition (Saneoka et al., 2004). As we know, nutrient elements are taken up by plant as solution, thus, water deficit indirectly causes less absorption of this element and consequently reduces plant growth and TB. Oweis et al. (2004) indicating grain yield and biomass loss of lens under drought conditions and they showed that above mentioned traits were improved as a result of irrigation.

The aim of this study was evaluation of effects of drought stress and nitrogen fertilizer on RWC, MSI, LCC, LA, stem length, stem dry weight and TB and to determine how much nitrogen is required to the plant especially in the low rate nitrogen fixation under water deficit conditions.

MATERIALS AND METHODS

In order to investigate the drought stress and nitrogen fertilizer effects on some traits of lens cultivar Gachsaran such as RWC, MSI, LCC, LA, stem length, stem dry weight and TB, a factorial experiment based on randomized complete block design as hydroponics was carried out with three university of mohaghegh Ardabili, Ardabil, Iran, in 2007. Factors were: drought levels (0, -0.6, 0.9 and -1.2 MPa using PEG) and nitrogen levels (0, 0.5 and 1.0 Mmol) stable aeration system for root respiration was used. Seeds were inoculated (infected) with RLV 73 which contained nitrogen fixator bacteria and then were sown in the sand. There after, seeding were transferred into 2 L pails involving nutrient solution. Photoperiod was adjusted as 16 h light and 8 h dark plants placed in the holes of the pails and their roots contacted with solution. This solution renewed each 12 days. Reduced water was recompensed daily with distilled water. Hoagland nutrient solution (Table 1) was used (Mehravaran, 2003).

Table 1: Elements and their rates used in the Hoagland solution
Image for - Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative 
        Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits 
        of Lentils (Lens culinaris L.) Under Hydroponics Conditions

After establishment and favorite growth of seedlings, nitrogen treatments were exerted. PEG treatment was done after words. At the end of treatment sampling was made. RWC was measured as follows (Rosales-Sernaa et al., 2004):

Image for - Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative 
        Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits 
        of Lentils (Lens culinaris L.) Under Hydroponics Conditions

FW, DW and TW were: fresh, dry and turgid weight, respectively. Sampling was done from newly-expanded leaves. Leaves immediately weighed and then immerged in distilled water for 5 h. Thereafter, turgid weight was obtained and finally, dry weight was measured after 24 h at the oven of 75°C. LCC was measured using chlorophyll meter (Conicaminalta-SPAD, Tokyo, Japan) (Rosales-Sernaa et al., 2004). Measurement was done at two staged: 1- After nitrogen treatment, 2- After drought stress treatment to calculation of LA. Some leaves were choose and their area was measured using leaf area meter (Scanman, USA), There after were dried at the oven of 75°C for 48 h finally, leaves were weighed and comparing leaf area and leaf weight, total leaf area for whole plant was gained. In order to measuring MSI, newly full expanded leaves were washed 3 timed with distilled water to removing surface electrolytes. In control treatment tubes 20 mL of distilled water and in stress one, 20 mL of PEG (M = 6000) were used. Samples were immersed in 40°C for 24 h and then there were washed three times. Then both tubes were filled with distilled water and samples were placed in, for nix 24 h until reaching temperature of the environment. Electrical conductivity (EC) was measured as Mm hoss-1. Samples were autoclaved for 25 min in 115°C and the second EC were measured MSI was calculated ad follows (Saneoka et al., 2004):

Image for - Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative 
        Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits 
        of Lentils (Lens culinaris L.) Under Hydroponics Conditions

where, IP is injury percent, T1, T2 is the EC of the stress treatment before and after autoclaving, respectively, C1, C2 is the EC of the control treatment before and after autoclaving, respectively. Length and dry weight of stem and TB were measured. Analysis of variances and mean comparisons were done using SAS software. Graphs were drawn by Excel program.

RESULTS AND DISCUSSION

RWC and MSI
Results showed that with increasing drought stress, RWC significantly (p<0.01) was increased and reached from 94.75% (control) to 76.74% (Table 2). Nitrogen had significant effect, as well and the most RWE was obtained at 0.5 Mmol nitrogen. Turkan et al. (2005) stated that water deficit obviously decreased RWC in bean and their findings is in accordance with present results. Also, MSI was decreased (p<0.05) under stress and reached to 73.09 from 81.39% in control (Table 2) but nitrogen had no significant effect on it. Confronting with drought causes to increase in gene transcriptions which oxidant cell wall lipids and consequently destroy the wall (Saneoka et al., 2004).

Table 2: Mean comparison of measured traits under drought stress and nitrogen
Image for - Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative 
        Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits 
        of Lentils (Lens culinaris L.) Under Hydroponics Conditions
*No. with same words in each column, have no significant differences to each other

Image for - Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative 
        Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits 
        of Lentils (Lens culinaris L.) Under Hydroponics Conditions
Fig. 1: Effects of different nitrogen levels on stem length under various root water potentials

Image for - Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative 
        Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits 
        of Lentils (Lens culinaris L.) Under Hydroponics Conditions
Fig. 2: Effects of different nitrogen levels on stem dry weight under various root water potentials

Length and Dry Weight of Stem and TB
Results showed that length and dry weight of stem and TB was decreased under drought stress so that, its rate was observed of 2.12 g plant -1. Length and dry weight of stem were measured of 32.14 cm and 0.50 g, respectively (Table 2).

Drought and nitrogen treatments has significant (p<0.01) effects on measured traits and had significant interaction effects on stem length (Fig. 1) and stem dry weight (Fig. 2) (p<0.01). Water has large effect on plant growth, biomass accumulation and nitrogen fixation and in case of water deficit, above mentioned aspects, decrease (Tomas et al., 2004). Athar and Johnson (1996) showed that reduction of water potential from -0.3 to -1.0 MPa, reduced biomass of 65%.

In this study, increase of nitrogen up to 0.5 Mmol, increased length and dry weight of stem and TB but in 1.0 Mmol nitrogen measured traits were decreased. This may because of either inactivation of nitrogen fixation in roots or insufficient values of nitrogen in absence of fixation.

Image for - Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative 
        Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits 
        of Lentils (Lens culinaris L.) Under Hydroponics Conditions
Fig. 3: Effects of different nitrogen levels on LCC under various root water potentials

Image for - Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative 
        Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits 
        of Lentils (Lens culinaris L.) Under Hydroponics Conditions
Fig. 4: Effect of different nitrogen levels on LCC after and before stress in various root water potentials

Image for - Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative 
        Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits 
        of Lentils (Lens culinaris L.) Under Hydroponics Conditions
Fig. 5: Effects of different nitrogen levels on LA in various root water potentials

LCC and LA
Both drought stress and nitrogen treatment had significant (p<0.05) on LCC and LA. Their interaction effects on LCC after drought stress (Fig. 3), LCC after and before drought stress (Fig. 4) and LA (Fig. 5) were significant (p<0.05).

The most difference between LCC before and after stress in severe drought conditions (-1.2 MPa) was obtained of 13.855 mg g-1 leaf dry weight, whereas, this value in control treatment was 1.877 mg g-1 leaf dry weight usually, water stress accelerates leaf senescence which has been seen in chickpea and lentil (Davies et al., 1999). The most LCC was got subjecting to 0.5 Mmol nitrogen under water stress (Table 3). Leaf production and expansion is very sensitive to available water and the reason is that cell division and growth need to water (Pagter et al., 2005).

Table 3: Mean comparisons of measured traits under drought stress and nitrogen levels
Image for - Evaluation of Water Stress and Nitrogen Fertilizer Effects on Relative 
        Water Content, Membrane Stability Index, Chlorophyll and Some Other Traits 
        of Lentils (Lens culinaris L.) Under Hydroponics Conditions
*No. with same words in each column, have no significant differences to each other

Brevedan and Egli (2003) reported that LCC and LA of soybean were decreased under water deficit RWC, LCC and stem biomass were decreased under the same conditions (Rosales-Sernaa et al., 2004).

CONCLUSIONS

Generally it was seen that whit increasing water deficit, all traits were decreased. The most RWC, LCC, LA stem nitrogen. So, it can be said that lentil has less demand than other legumes and it seems that 0.5 Mmol nitrogen is sufficient for plant growth and excessive values mighty have unflavored effects because of reducing nitrogen fixation by plant and being insufficient for total plant requirement for this element. Therefore, adequate rates of nitrogen under water stress conditions noticing fixation capability, is recommended.

ACKNOWLEDGMENTS

This study was supported by the Central Laboratory of Agricultural Faculty, University of Mohaghegh Ardabili. Valuable experimental support by Aziz Jamaati-e-Somarin and Asad golizadeh is greatly appreciated. This study was extracted from M.Sc. Thesis of Mitra Salehpour.

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