Legume Research

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Research Article

Legume Research, volume 44 issue 4 (april 2021) : 413-418

PGRs and nutrient consortium effect on water relations, photosynthesis, catalase enzyme and yield of blackgram under salinity stress

R. Sivakumar1,*, S. Jaya Priya1
1Regional Research Station, Tamil Nadu Agricultural University, Paiyur-635 112, Tamil Nadu, India.

  • Submitted09-01-2019|

  • Accepted15-05-2019|

  • First Online 14-08-2019|

  • doi 10.18805/LR-4118

Cite article:- Sivakumar R., Priya Jaya S. (2019). PGRs and nutrient consortium effect on water relations, photosynthesis, catalase enzyme and yield of blackgram under salinity stress . Legume Research. 44(4): 413-418. doi: 10.18805/LR-4118.

ABSTRACT

An experiment was conducted to study the effect of plant growth regulators namely jasmonic acid (50 µM), brassinolide (0.5 ppm), salicylic acid (100 ppm), ascorbic acid (100 ppm), gibberellic acid (10 ppm), benzyl amino purine (5 ppm) and nutrient consortium (K2SO4 (0.5%) + FeSO4 (0.5%) + Borax (0.3%) and TNAU Pulse Wonder (1%) on blackgram (Vigna mungo L.) variety TNAU (Blackgram) CO  6 under salinity condition. The salinity was imposed by 125 mM NaCl concentration in pot culture. Plant growth regulators and nutrient consortium were used as foliar spray at 20 and 40 days after sowing except jasmonic acid, which used as seed soaking. Under salinity stress, water potential, relative water content (RWC), photosynthetic rate, chlorophyll index and yield were significantly reduced as compared to control. Among the treatments, brassinolide and TNAU Pulse Wonder showed supremacy to enhance water potential, RWC and yield compared to other treatments. Salicylic acid performed better to stabilize the photosynthetic rate under salinity followed by TNAU Pulse Wonder. Benzyl amino purine had higher chlorophyll index than other treatments. Increased catalase activity was observed in brassinolide treated plants followed by salicylic acid and ascorbic acid under salinity.

INTRODUCTION

Various environmental factors like salinity, drought, heavy metal and temperature adversely affect plant growth and development and finally the biological yield of the crops. Salinity is one of the most important abiotic stresses, limiting crop production in arid and semi-arid regions, where soil salt content is naturally high and precipitation may be insufficient for leaching (Zhao et al., 2007).
 
Soil salinity reduces the productivity of many agricultural crops, which are particularly sensitive throughout the ontogeny of the plant (Machado and Serralheiro, 2017). Sonia Rani et al., (2019) found that the soil salinity affects plant growth by creating water and ionic imbalance in the plant due to the presence of toxic ions. Salt-affected plants exhibit stunted growth and have darker leaf colour. Salinity inhibits the physiological and biochemical processes basis for imbalance in metabolism and enhances the production of reactive oxygen species which disturbs the plant growth and development ultimately yield reduction (Zorb et al., 2019).
 
Blackgram is one of the important pulse crops which have its origin in India. It is favorable short duration pulse crop and well suited in all seasons either as sole or as intercrop or fallow crop. India is the world’s largest producer as well as consumer of blackgram.  It has high nutritive value and consist high protein content (26%), vitamins and minerals. Munns (2002) reported that the blackgram is very sensitive to salinity in early stage of growth and affects the ability to utilize water and cause a reduction in growth rate as well as plant metabolic processes ultimately yield.
 
Plant growth regulators (PGRs) and nutrients have been used to promote plant growth and development of plants under various stress conditions. Ali et al., (2007) reported that brassinosteroids are a class of polyhydroxy steroidal lactones that play essential roles in plant growth and development and amelioration of the salt stress in different plant species. Foliar application of salicylic acid increased the photosynthetic rate in corn and soybean under salt stress (Khan et al., 2003). Kaydan et al., (2007) found that the application of ascorbic acid alleviates the destructive effects of salinity on water potential and photosynthetic pigments in wheat seedlings under salinity stress. Cytokinins could increase salt tolerance in wheat plants by interacting with other plant hormones, especially auxins and ABA (Iqbal et al., 2006). Plant growth and salt tolerance were sharply reduced when exposed to a combination of salt stress and potassium deficiency stress. Degl’Innocentiet_al(2009) reported that the potassium deficiency significantly increased the negative effects of salt on photosynthesis in barley and was accompanied by an increase in salt sensitivity. Tamil Nadu Agricultural University (TNAU) Pulse Wonder is a nutrient and PGR consortium developed by Tamil Nadu Agricultural University which contains essential nutrients and PGRs required for pulse crops like greengram and blackgram. Hence, the experiment was conducted to study the performances of different plant growth regulators and nutrient consortium on salt tolerance by assessing physiological traits like water potential, RWC, photosynthetic rate, chlorophyll index, catalase enzyme activity and yield of blackgram under salinity condition.

MATERIALS AND METHODS

A pot culture experiment was conducted in glass house at Department of Crop Physiology, TNAU, Coimbatore during 2016-17 in blackgram. Red sandy soil mixture (red soil, sand and vermicompost in the ratio of 2:1:1 respectively) was used for pot culture experiment.
 
Blackgram is a direct seed sown crop, hence seeds were sown in a pot then the salt solution was poured on the soil. Salinity of 125 mM NaCl was standardized for blackgram in the previous experiment in which 50% yield reduction has been observed. The same level of saline water was added to the soil as in the case of farmers’ field in which they are irrigating the field with saline water. Hence, salinity (125 mM NaCl) was imposed from sowing onwards till the end of the harvest. Crop was applied with recommended dose of fertilizers (25 kg N + 50 kg P2O5 + 25 kg K2O per hectare) and other cultivation operations including plant protection measures were carried out as per recommended package of practices of Tamil Nadu Agricultural University, Coimbatore. The experiment was laid out in completely randomized block design with ten treatments and three replications.
 
Plant growth regulators like gibberellic acid (10 ppm), brassinolide (0.5 ppm), salicylic acid (100 ppm), ascorbic acid (100 ppm), benzyl amino purine (5 ppm) and nutrient consortium like K2SO4 (0.5%) + FeSO4 (0.5%) + Borax (0.3%) and TNAU Pulse Wonder (1%) were used as foliar spray at 20 and 40 DAS. In this study, 50 µM jasmonic acid was used as seed soaking PGR. The seeds were soaked in jasmonic acid solution for four hours and then shade dried. The observations like water relations, gas exchange and catalase activity were recorded at 30 and 50 DAS. The yield parameters were estimated after harvest.
 
Water relation parameters
 
Measurement of leaf water potential
 
Leaf water potential was measured by using an instrument Leaf Water Potential Meter (ARIMAD 3000) and expressed as MPa.
 
Estimation of relative water content
 
Relative water content was estimated according to Barrs and Weatherly (1962) and calculated by using following formula and expressed as per cent.
 
 
 
Gas exchange parameter-measurement of photosynthetic rate
 
Photosynthetic rate measurement was performed by using Portable Photosynthesis System (PPS) (Model LI-6400 of LICOR inc., Lincoln, Nebraska, USA) equipped with a halogen lamp (6400-02B LED) positioned on the cuvette.
 
Third leaf from top was used for the measurements with replicated thrice. Leaf was inserted in a 3 cm2 leaf chamber and PPFD at 1200 μmol photons m-2 s-1 and relative humidity (50-55%) were set. The readings were taken between 9 am to 11.30 am and the value is expressed as μmol CO2 m-2 s-1.
 
Chlorophyll Index (SPAD value)
 
SPAD readings were recorded by using chlorophyll meter (SPAD 502) designed by the soil plant analytical development (SPAD) section, Minolta, Japan. The Minolta SPAD-502 measures chlorophyll content as ratio of transmittance of light at wavelength of 650 nm and 940 nm. Five readings were taken from each replication and the average value was computed using the method described by Monje and Bugbee (1992).
 
Catalase activity
 
Catalase activity was determined by following titration method using potassium permanganate (Gopalachari, 1963) expressed as μg H2O2 g-1 FW min-1. 500 mg of leaf sample was macerated with 10 ml of phosphate buffer and centrifuged the content at 3000 rpm for 10 minutes. 1 ml of supernatant was taken to 5 different beakers and 5 ml of 1.5 % sodium perborate and 1.5 ml of phosphate buffer were added. 10 ml of 2 N sulphuric acid was added to each beaker at the time interval of 1 minute, 2 minutes, 3 minutes and 4 minutes after adding enzyme extract in first four beakers respectively. In the final beaker, 10 ml of sulphuric acid was added before addition of the enzyme extract. This fifth beaker was kept as blank for comparison. The contents in the beaker were titrated against 0.05 N KMnO4. Development of pink color which persists for 30 seconds is the end point. The volume of KMnO4 consumed was noted. One ml of KMnO4 is equal to 0.85 μg of H2O2. The activity of the enzyme is expressed as μg of H2O2 g-1 FW min-1.
 
Yield
 
The total weight of grains harvested from each plant of all picking was added and average yield per plant was worked out and expressed in gram per plant. The data on various parameters were analyzed statistically as per the procedure given by Gomez and Gomez (1984).

RESULTS AND DISCUSSION

Decreasing trend of leaf water potential was observed from 30 DAS to 50 DAS. In the present experiment, leaf water potential was decreased up to 44 per cent due to salinity when compared to normal condition. At 30 DAS, the maximum leaf water potential was recorded in TNAU Pulse Wonder followed by brassinolide and salicylic acid. Brassinolide recorded the maximum leaf water potential followed by TNAU Pulse Wonder, salicylic acid and ascorbic acid at 50 DAS (Table 1). Hayat et al., (2012) found that brassinosteroid had a favorable impact on the leaf water potential of the plants under abiotic stress condition in tomato. The positive role of TNAU Pulse Wonder on water potential might be due to act as an osmoticum, as it contains compatible osmolyte nutrients.
 

Table 1: Effect of PGRs and nutrient consortium on physiological traits in blackgram under salinity.


 
Relative water content is considered as the index which indicates the ability of the plants to retain tissue water status under stress and the plants retaining more tissue water are expected to perform better under any stress condition. In the present experiment, high relative water content values of 85.7 per cent observed in absolute control and low relative water content was observed under salinity. The results exhibited the significant difference in all the treatments. Among the treatments, brassinolide recorded higher relative water content which is on par with salicylic acid followed by ascorbic acid and TNAU Pulse Wonder at 50 DAS. Reduction of RWC under salinity condition might be due to reduced water uptake by high osmotic pressure of the soil.
 
Significant reduction in relative water content was noticed in blackgram (25%) under salinity stress compared to absolute control. In this present study, brassinolide treatment increases the relative water content up to 12.38 per cent followed by salicylic acid (11.89%) compared to control. The results are also coincides with the findings of Gograj Jat et al., (2012) who reported that use of brassinolide up to 1.0 ppm increased the relative water content, photosynthetic rate and transpiration rate significantly at flowering stage of cluster bean cultivars under salinity stress. Aldesuquy (2014) reported that sodium salicylate appeared to regain the leaf turgidity under salinity condition by inducing additional decreases in transpiration rate through more reduction in stomatal pore.
 
Significant difference was noticed in all the treatments with respect to photosynthetic rate under saline condition. The higher photosynthetic rate was noticed in absolute control and the lower photosynthetic rate noticed in control at 30 DAS. Among the treatments, brassinolide recorded the higher photosynthetic rate which is on par with TNAU Pulse Wonder followed by ascorbic acid and salicylic acid. However at 50 DAS, salicylic acid registered higher photosynthetic rate which is on par with TNAU Pulse Wonder followed by brassinolide. In this study, photosynthetic rate was increased up to 42.31 in brassinolide treatment followed by TNAU Pulse Wonder (35.01%) over control (Table 1). The positive effect of TNAU Pulse Wonder is might be due it contains all essential nutrients and plant growth regulators required for blackgram, which are enhanced the photosynthetic rate. Kanmani et al., (2017) found that the foliar spray of 1 ppm brassinolide increased the photosynthetic rate and chlorophyll fluorescence under salinity stress in rice. Hayat et al., (2008) reported that the application of salicylic acid enhanced the chlorophyll content and photosynthetic rate and also neutralized the negative effect of the stress in tomato.
 
Chlorophyll index in terms of SPAD value was measured at different growth stages of plants and treatments spray showed significant differences. The higher SPAD value was registered in absolute control and lower value registered in control at 30 DAS and decreased at 50 DAS. Among the treatments, higher chlorophyll index was recorded in benzyl amino purine which is on par with brassinolide followed by TNAU Pulse Wonder. At 50 DAS, benzyl amino purine recorded higher chlorophyll index which is on par with TNAU Pulse Wonder followed by nutrient mixture and brassinolide. An inverse relationship between salinity and photosynthetic pigment content was observed earlier. When NaCl concentration increased in the growth medium, chlorophyll content decreased up to 52% in high-yielding genotype of Phaseolus vulgaris (Taibia et al., 2016).
 
 
The reduction of chlorophyll index and pigment might have been due to degradation of chlorophyll by increased chlorophyllase activity and generation of reactive oxygen species under salinity. Salt induced osmotic stress as well as sodium toxicity trigger to the formation of reactive oxygen species, such as superoxide, hydrogen peroxide, hydroxyl radical and singlet oxygen, which can damage mitochondria and chloroplast by disrupting cellular structure (Murakeozy et al., 2003).
 
The enhancement of chlorophyll index by benzyl amino purine might be due to act as anti-senescence hormone which protect the chlorophyll degradation under salinity. Wu et al., (2012) concluded that benzyl adenine could alleviate the detrimental effects of salt stress on plant growth by increasing chlorophyll, photosynthetic efficiency and enhancing antioxidant enzyme systems in egg plant. The chlorophyll index value was reduced up to 38 per cent  due to salinity and improved by benzyl amino purine (28.6%) followed by TNAU Pulse Wonder (21.75%).
 
The increment of chlorophyll index by TNAU Pulse Wonder over control under salinity might be due to increased synthesis of chlorophyll through catalytic role as it contains magnesium, which is important for chlorophyll assembly. The positive effect of brassinolide on chlorophyll protection under salinity might be due to involvement in membrane integrity by enhancing the level of the antioxidant system (Arora et al., 2008). Hayat et al., (2010) reported that plants exposed to high NaCl exhibited a significant decline in growth, photosynthetic parameters and a minimum quantum yield of PS II. However, the treatment with homobrassinolide, detoxified the stress generated by NaCl and significantly improved the values for the above parameters. The present investigation was supported by earlier findings.
 
In present study, the catalase activity assessed in the leaves of blackgram under salinity stress condition revealed a significant difference in the activity at 30 and 50 DAS.  The absolute control recorded the minimum catalase activity and control recorded the maximum value in 30 and 50 DAS. Hence, it was showed that the catalase activity increased under stress condition. The PGRs and nutrient treatments showed increased enzyme activity compared to control. Among the treatments, brassinolide recorded higher enzyme activity followed by salicylic acid and ascorbic acid. Significant differences were observed in all the treatments with respects to catalase activity under saline condition. At 50 DAS, higher enzyme activity was found in brassinolide followed by ascorbic acid which is on par with salicylic acid. The reactive oxygen species generation in plants may be responsible for most of the damage to cellular components. The enzyme catalase, peroxidase and super oxide dismutase involved in the detoxification of active oxygen species, especially hydrogen peroxide (Yamazaki et al., 2003). This increased activity of these enzyme act as control system and thus, provide protection from oxidative stress, which otherwise could cause membrane peroxidation, damage to cell organelles, and inhibit photosynthetic and other enzyme activities. Higher activity of catalase decreased H2O2 level in cell and increases the stability of membranes and CO2 fixation because several enzymes of the Calvin cycle within chloroplast are extremely sensitive to H2O2. A high level of H2O2 directly inhibits CO2 fixation.
 
Application of plant growth regulators and nutrient consortium increased the activity of catalase enzyme compared to untreated plants under salinity. In this present investigation, brassinolide treatment increased the catalase activity up to 37.46 percent followed by salicylic acid (35.09%) over control (Fig 1).
 

Fig 1: Effect of PGRs and nutrient consortium on catalase activity in black gram under salinity.


 
This result was supported by Mohammad et al., (2011) who studied with wheat (Triticum aestivum L.) cultivars and found that the treatment of salt stressed plants with homobrassinolide enhanced the enzymes activities of nitrate reductase, catalase, peroxidase, and superoxide dismutase. The application of brassinosteroids increases the accumulation of proline and enhances activities of antioxidant enzymes in salt stressed Vigna radiata (Hayat et al., 2010).
 
Comparing the treatments, absolute control recorded the highest grain yield of 10.30 g plant-1 and control recorded the least value of 7.18 g plant-1. Significant difference was recorded with treatments with respects to grain yield under saline condition. Among the PGRs and nutrients, brassinolide recorded the maximum grain yield followed the salicylic acid which is on par with TNAU Pulse Wonder (Table 2). 
 

Table 2: Effect of PGRs and nutrient consortium on yield in blackgram under salinity.


 
The foliar spraying of plant growth regulators and nutrients might have exploited favorably for indeterminate crop for continuous translocation of photosynthates to economic part and ultimately yield increment. Apart from this, delayed senescence may also be attributed for the increase in yield. The production of higher seed yield due to PGRs may be attributed to the fact that plants treated with growth regulators remained physiologically more active to build up sufficient food reserves for developing flowers and seeds.
 
Earlier studies, in the case of green gram also, foliar application of brassinolide and salicylic acid could be beneficial for improving yield and nutritional quality of green gram (Bera et al., 2008). Hossain et al., (2015) reported that the foliar application of salicylic acid at 200 ppm under different salinity conditions had the positive effects related to mitigation of salinity stress effect in chickpea.
 
Similar result was found in Jeyakumar et al., (2008) who reported that application of 125 ppm salicylic acid to blackgram plants increased seed yield. The spray of homobrassinolide to the foliage of Brassica juncea plants enhanced the growth, nucleic acid content and seed yield under NaCl condition (Hayat et al., 2006). Jyoti Verma et al., (2012) evaluated the effect of brassinolides on physiological traits in Cicer arietinum L. and reported that 1 ppm brassinolide significantly increased the plant height, number of pods per plant, biological yield, seed yield per plant, number of seeds per pod under stress condition. Vazquez et al., (2019) stated that the brassinosteroids stimulated salt stress tolerance in plants is associated with decrease of oxidative damage created by salinity and the interaction between brassinosteroids and other plant hormones like salicylic acid. Foliar application of brassinolide to rice young plants increased the tolerance by showing increased shoot dry weight under saline conditions.
 
With respect to TNAU Pulse Wonder, the possible reason for improvement in yield might be due to nature of this crop booster with combination of nutrients and growth regulators for pulse resulted in decreased flower shedding and improvement in crop tolerance to abiotic and biotic stress.

CONCLUSION

Our results showed that brassinolide, salicylic acid and TNAU Pulse Wonder have a greater role in alleviation of salinity effect in blackgram. Under salinity stress condition, brassinolide and TNAU Pulse Wonder had a positive role on water potential and relative water content and protects the plant from salinity damage. Salicylic acid and TNAU Pulse Wonder have also positive impact in the case of photosynthetic rate and catalase activity. Benzyl amino purine has a supremacy act to protect the plant under salinity through protecting chlorophyll.

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